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Brauer Notebook

Captains’ Log. iGEM 2014. Team NitroGENIUS. The Brauer Group.

Day 1. 6/2/2014 (Monday)

● Plan to do: design primers/plasmids

● Got done: started primer/plasmid design

Day 2. 6/3/2014 (Tuesday)

● Plan to do: Create and work on presentation

● Got done: Monsanto Presentation

Day 3. 6/4/2014 (Wednesday)

● Plan to do: Monsanto Visit

● Got done: Monsanto Visit

Day 4. 6/5/014 (Thursday)

● Plan to do: More plasmid editing/primer design. Edit website. Plan out video series.

● Got done: Finished pLight plasmid design and primers, sent to Cheryl for verification. Typed up brainstorming for video sessions, contacted WUTV about using equipment, contacted WashU Assembly Series director for permission to allow a Monsanto scientist to give a lecture during the Fall 2014 semester

Day 5. 6/6/2014 (Friday)

● Plan to do: Get iGEM safety forms filled out. Go over primer/plasmid design. Order primers. Get keys/access to Brauer Hall. Design a timeline for the summer.

● Got done: Finished design of plasmid PBJ001. Finished all (including sequencing) primer design. Take another look on Sunday, 6/8/2014.

Name	Primer
EYFP-> ColE1 _f	5’ tttttttgctcttcaaatGGGATAACGCAGGAAAGAAC 3’
EYFP-> ColE1 _r	5’ tttttttgctcttcagtgCTGAAGGCTGACAAAATCTATACTG 3’
Pcpcg2 -> Ptet_f	5’ tttttttgctcttcaTTCCCAGTCACGACGTTG 3’
Pcpcg2 -> Ptet_r	5’ tttttttgctcttcaTCGGTCAGTGCGTCCTG 3’
TetR_f	5’tttttttgctcttcaCACCTACAACAAAGCTCTCATC 3’
TetR_r	5’tttttttgctcttcagaaGGCTAGCTCAGTCCTAGGTACAG 3’
EYFP_f	5’ tttttttgctcttcaattCTTGTGCAATGTAACATCAGAG 3’
EYFP_r	5’ tttttttgctcttcacgaGTGGAATTGTGAGCGGATAAC 3’
Seq1_f	5’ ACAAATCCGCCGCCCTAG 3’
Seq1_r	5’ GACTTGAGCGTCGATTTTTGTG 3’
Seq2_f	5’ GGCTTTTTTACCGGCTAGTTTC 3’
Seq2_r	5’ GATTTTCTCCCCCTTTTCTTC 3’

Day 6. 6/9/2014 (Monday)

● Plan to do: Get iGEM safety forms filled out. Order primers. Get keys/access to Brauer Hall. Design a timeline for the summer. Complete lab safety forms for WashU. Design primers/plasmid map for positive/negative control.

● Got done: Ordered primers (should arrive by Wednesday at the latest). Got keys/access to Brauer Hall. Completed lab safety forms for WashU.

Day 7. 6/10/2014 (Tuesday)

● Plan to do: Finish design of pos/neg control plasmids. Print Brauer lab safety form. Finalize proposal and send to Penn State iGEM contact along with project presentation. Design light experiment.

● Got done: Designed primers for the positive and negative control plasmids. Printed safety form. Sent Penn State iGEM project info. PCR, ran visualization, and extracted pieces.

Name	Primer
PJT122_modNeg_minuslacZ1_Forward	GGTTTTCCCAGTCACGACG
PJT122_modNeg_minuslacZ1_Reverse	CTGGAGCCCGTCAGTATCG
PJT122_modNeg_minuscph8_2_Forward	GCAGGGCATGACAAAAGAAG
PJT122_modNeg_minuscph8_2_Reverse	CTGAGTTGTACGGTGGTGGC
PJT122_modNeg_seq_cph8_Foward	CAACGTGCCAACCAGTTTTAG
PJT122_modNeg_seq_cph8_Reverse	GAACGACCTACACCGAACTG
PJT122_modNeg_seq_lacZ_Forward	CAAAAACGCCGTCAATTTG
PJT122_modNeg_seq_lacZ_Reverse	GCTTTACACCGTTGATATTGC
pSL2264_EYFP_pos_Foward	tttttttgctcttcaCGTGGCTTTGTTGAATAAATC
pSL2264_EYFP_pos_Reverse	tttttttgctcttcatcaAGCGGATAACAATTTCACACATAC
pTetR100_pos_Forward	tttttttgctcttcaTGAGCTAGCACTGTACCTAGGAC
pTetR100_pos_Reverse	tttttttgctcttcaacgCTGAAGTGGCGGAACTGC
pTetEYFP03_seq_Forward	GGTCATGGTAAAGGCAATGG
pTetEYFP03_seq_Reverse	TATGGCAATGGCACAATTAGATG

Day 8. 6/11/2014 (Wednesday)

● Plan to do: Moon Lab Group meeting at 3PM. Work on finishing Gel DNA extraction, Dpn1, Purification, then One-Pot, then finally Transformation and plating (2 LB/cm plates). [Realization! We discovered today that in order for the ccaS/ccaR light system to function correctly, we need the PCC6803 genes ho1 and pcyA which are needed to heme into PCB (a chromophore) via a two step oxidation/reduction process. Ray is working on this aspect and we are planning to help/take over this part of his work. Our plan is to still transform plasmid PBJ and send it off for sequencing to make sure we have our designed plasmid. The next step would be to co-transform the two plasmids, one containing the ho1 and pcyA genes and the plasmid PBJ into E. coli and after sending off for sequencing, test the light promoter system. ]

● Got done: Finished Gel DNA extraction and measured concentration using nanoquant tecan. Did not need to do Dpn1 nor purification. Put in one-pot after 12 noon. Transformed and incubated O/N. PCR for Ray’s Biobrick plasmid for ho1 and pcyA parts.

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
TetR	143.1	EYFP	128.4
Col A	12.3	Col B	5.8
Col D	12.9	Col E	2.8
G2 A	26.2	G2 B	26.2
G2 C	45.4	G2 D	26.6
G2 E	15.3	G2 F	77.7

Day 9. 6/12/2014 (Thursday)

● Plan to do: Gel extract, one-pot for Ray’s plasmid then transform and plate. pick PBJ002 colonies, start O/N liquid cultures. Work on negative and pos. control once primers arrive (check around noon). Also, grow a culture of Ray’s BbaP DNA (AmpR) as well as Cheryl’s pTet 100-T (where we get the TetR from)

Plan out seq PCR for PBJ002.

● Got done: Gel extract and one pot for Ray’s plasmid. Re-ran PCR for Rays BbaP (SpeC_AraC_Bba) plasmid. Gel extract on new BbaP part. Did not yield bands- only saw a smear. Received primers for neg and pos controls- mixed appropriate primer concentrations. Cheryl started pTet liquid culture. Started BbaP liquid cultures. Electroporated original Bbap. Plated on 2 specR plates (100 uL) and (the rest). put into incubator.

Planned out seq PCR for PBJ002.

Day 10. 6/3/2014 (Friday)

● Plan to do: meet at 9:15 in the ICARES to set up video conference with the Penn State.

1. PBJ002: Miniprep -> PCR with sequencing primers before ICARES meeting

miniprep pTet and BbaP we made liquid cultures for yesterday. miniprep this along with the PBJ miniprep.

2. pour gel for visual confirmation

...ICARES meeting… then...

3. take PCR product and run a gel (10 uL DNA, 2uL 6X dye) to check. 30 minutes.

Observe which PCR product in gel (which lane from tube) is of the right length.

~~Purify PCR product.~~

Send miniprepped plasmid ~~AND~~ ~~purified PCR~~ off for sequencing of PBJ002.

4. work on both the Positive and negative controls.

5. Pick colonies and start O/N culture for Spec_AraC_Bba.

● Got done: From the above mentioned steps: Steps 1- 5.

○ For the Pos and Neg controls, we did PCR -> nanoquant measure after purification step (reference pos/neg control step documents) TetR backbone not present. Re-run tomorrow.

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
PBJ 02 #1	103.9	PBJ 02 #2	90.1
PBJ 03 #3	137.5	PBJ 02 #4	76.9
Ptet	60.4	BbaP	62.2
(-) lacZ #2	103.9	(-) lacZ #6	90.1
(-) lacZ #3	137.5	(-) lacZ #7	76.9
(-) lacZ #5	60.4	(-) lacZ #8	62.2
EYFP	34.6

Day 11. 6/14/2014 (Saturday)

● Plan to do:

○ Miniprep -> Visualize seq PCR product of Spec_AraC_Bba

■ could not run seq PCR on this because seq primers do not work. Re-design sequencing primers and order them.

○ Re-run PCR for TetR backbone. Failed yesterday.

○ ~~One pot Pos control~~

○ One pot Neg control

...Later...

○ Transform/plate neg control

○ ~~Transform/plate pos control~~

● Got done:

○ Miniprepped and measured concentration on SAB product.

■ Project SAB is on hold until primers arrive.

○ Re ran PCR on the TetR backbone.

○ sent new redesigned sequencing primers for SAB to Ray for ordering.

○ Gel Extraction/Recovery on TetR backbone PCR product

○ Dpn1 for TetR backbone

○ TetR purification and measurement= 4.4 ng/uL

■ Project Positive Control is on hold until we can re-evaluate the primers and find the error/form a new plan of approach.

■ Correction: re-did PCR for TetR backbone again. (was the reason we couldn’t get it to work before because we used the wrong plasmid?)

● gel only yielded bands indicating primers.

○ One pot Negative control (-) lacZ

○ Transform/plated Negative control (-) lacZ

Day 12. 6/15/2014 (Sunday)

● Plan to do:

○ TetR backbone for positive control run from PCR- >Transformation/Plating

○ Pick Neg Control colonies.

■ Start liquid cultures for O/N

● Got done:

○ (+) TetR backbone from TetR-100T miniprepped plasmid: PCR -> DNA purification

○ Picked Neg Control colonies.

■ Started liquid cultures for O/N

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
TetR Backbone A	148.4	E	75.9
B	123.5	F	76.7
C	109.1	G	78.0
D	29.3	H	74.3

Day 13. 6/16/2014 (Monday)

● Plan to do:

○ Negative control: freeze and miniprep -> Send to Medical School for sequencing

○ Positive control: One-pot -> Transform/plate

● Got done:

○ Freeze and miniprep negative control

○ Negative Control: Ran Sequencing PCR for (-) lacZ

■ Visualization gel (smear)

■ Rerun sequencing PCR with new settings- failure

Redux starting with DPN1 (-) lacZ #5

■ Purify

■ Nanoquant

■ Blunt end ligation

■ Transformation/Electroporation and plating on cm plates

○ Positive Control (EYFP + TetR backbone) Run One Pot

■ Transformed

■ plated 100uL, rest at 8:05pm

■ Incubate o/n

Day 14. 6/17/2014 (Tuesday)

● Plan to do: Pick both Positive and Negative Control colonies

○ Positive: kan antibiotic

○ Negative: cm antibiotic

■ no colonies seen as of 10 AM

■ colonies seen at 12 noon!

● Got done:

○ redesigned new PBJ003 plasmid map and designed/verified new sequencing primers. Sent them to Cheryl for verification. SAB primers have not yet arrived. Liquid cultures into the incubator by 9PM

Day 15. 6/18/14 (Wednesday) -Worldly Wednesday!

● Plan to do:

○ Get both Pos and Neg frozen and miniprepped -> send off for sequencing

■ concurrently with SAB?

○ Go to Worldy Wednesday Lunch hosted by Andrew Ng

○ Work on SAB?

● Got done:

○ Froze both Pos and Neg and miniprepped.

■ Pos process went to sequencing.

■ Negative control: start at seq PCR.

○ Ran SAB SeqPCR.

■ negative results

○ Benjamin Colony PCR negative control (- lacZ) from old plates

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
Pos #1	54.1	Neg #1	92.5
Pos #2	163.4	Neg #2	73.1
Pos #3	63.7	Neg #3	78.8
Pos #4	109.5	Neg #4	126.5

Day 16. 6/19/14 (Thursday)

● Plan to do:

● Got done:

○ Re-designed the SEQ primers for SAB.

○ CPCR for (-)lacZ

■ move on to (-)cph8

○ begin work on PBJ003

Name	Primer
TetR_forward_new	tttttttgctcttcacacTCTCTACTAGAGTCACACTGGCTC
EYFP_forward_new	tttttttgctcttcaattGGGAGCACTGTATTCACAGCGCTCCCtttaccggtctcaattcgcag
Spec2_Seq_For	CGAGATAACCGTTGGCCTC
AraC2_Seq_For	gttatccaataataactctgatagtgctag
Bba2_Seq_Rev	cacttcactgacaccctcatc

Day 17. 6/20/14 (Friday)

● Plan to do:

● Got done:

○ PCR and gel extracted correct EYFP

○ working on getting piece Ori PCR -> gel extract and setting up one-pot for PBJ003

○ (-) cph8 took all the way through DNA clean & concentrate and added wrong wash buffer

■ 5µl purified DNA remaining in -20 (inside cph8 box)

Day 18. 6/21/14 (Saturday)

● Plan to do:

● Got done:

○ Negative Control:DPN1, purify, nanoquant measure, blunt end ligation (BEL) and transform/plate on cm

○ PBJ003: PCR -> DNA Recover and Nanoquant measure. Digestion/Ligation (One-Pot) all pieces. Transformation/Plating on cm

Day 19. 6/22/14 (Sunday)

● Plan to do: Pick colonies start liquid colonies for -cph8, pbj003

● Got done: Pick colonies start liquid colonies for -cph8, pbj003

Day 20. 6/23/14 (Monday)

● Plan to do: Freeze, miniprep, sequencing pcr, visualization gel, send for sequencing

● Got done: negative control, pbj003: Freeze, miniprep, sequencing pcr, visualization gel, send for sequencing

○ SAB: Sequencing PCR, visualization gel

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
Neg #1	128	PBJ003 #1	146.3
Neg #2	81.2	PBJ003 #2	86.9
Neg #3	47.8	PBJ003 #3	69.4
Neg #4	50.7	PBJ003 #4	95.2

Day 21. 6/24/14 (Tuesday)

● Plan to do: design experimental methods and procedures, design around hybrid promoter, find lacI resistant strain

● Got done:

○ did modified SEQPCR on SAB and ran visualization gel- confirmed S and B present, A not present

○ Ran PCR on part AraC/p15a ( part A), extracted and DNA recovered along with nanoquant measurements.

○ S,A,B Digestion

○ S+A+B Ligation into SAB2

○ Transformation and Plating of SAB2 (failed)

Day 22. 6/25/14 (Worldly Wednesday)

● Plan to do: Transform SAB2 C and D. Check sequencing results for the negative control and PBJ003 if they come in today.

● Got done: Verfied sequences for some positive control #4, some PBJ003. Sent T2 and E4 of PBJ003 in for sequencing confirmation, send in (-) lacZ sequencing on (-)cph8 plasmid, transformed and plated SAB2 C and D. Design primers for hybrid promoter

Day 23. 6/26/14 (Thursday)

● Plan to do: Pick and grow colonies for liquid culture

● Got done: Grew liquid Colonies for SAB2. Work on website. Look up experimental procedures

Day 24. 6/27/14 (Friday)

● Plan to do: Freeze and miniprep SAB2; Sequencing PCR, Send for sequencing

● Got done: Freeze and miniprep SAB2; Sequencing PCR, PCR fail, will re run with a gradient. Verified completely PBJ003 #2, #4. (-) lacZ funky. Received primers for hybrid promoter

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
SAB #1	86.6	SAB #2	97.1
SAB #3	79.1	SAB #4	65.8

Day 25. 6/28/14 ( Saturday)

● Plan to do: Run Sequencing PCR (gradient) for SAB2C #2. Send out purified PCR product for Sequencing. Set up experimental procedures, write out experiment protocol.

● Got done: Ran sequencing PCR with a gradient for SAB2 C#2. Verified negative control completely finished.

Day 26. 6/29 /14 ( Sunday)

● Plan to do: Run SEQPCR (gradient) for SAB2C #2 and visualize.

● Got done: Ran SEQPCR for SAB2C #1. Same results on from 6/28/14. Redesigned primers for new plasmid (not using SAB anymore) called pHOP

Day 27. 6/30 /14 ( Monday)

● Plan to do:

● Got done: picked ice mix from Ray’s pSB1C3-BBa_M30109 frozen stock. Preparing for miniprep tomorrow morning (0800).

Day 28. 7/1 /14 ( Tuesday)

● Plan to do: Miniprep pSB1C3-BBa_M30109. Do a digestion and gel visualization step for pSB1C3-BBa_M30109 to see if its legit. Start PCR and steps for PBJ_Hybrid (PBJ003 + hybrid promoter)

● Got done: Saw nothing in the first digestion, could not re-run digestion due to no enzyme left. PCR, DPN1, Clean, BEL of PBJ_Hyb_001, transform, plate

Name	Primer
PBJ_hyb_for	5’ cattatacgagccggatgattaattgtcaa ttt CATTAAAAATGCGATCCTAAC 3’
PBJ_hyb_rev	5’ tgtggaattgtgagcggataacaatttcacaca ATGATAGATCCCGTCGTTTTAC 3’

PBJ_hyb_seq_for	5’ GCGACTTGATGCTCTTGATC 3’
PBJ_hyb_seq_rev	5’ CGAGGTCATATTCCAGTCG 3’

Day 29. 7/2/14 (Worldly Wednesday)

● Plan to do: make LB/cm plates (follow protocol), look at plated cultures and wrap when necessary. pHOP primers should arrive today so we can get started working on that

● Got done: Pulled plates (2) from incubator, wrapped them and put into 4 fridge. Picked colonies for PBJ_hyb_001. started pHOP procedures.

Day 30. 7/3/14 (Thursday)

● Plan to do: Miniprep -> send off for sequencing for PBJ_hyb_001. One pot then Transform and plate pHOP

● Got done: Miniprep-> visualize for PBJ_hyb_001. Plan on preparing for sequencing on Sunday. One pot, transformed and plated pHOP.

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
PBJ_hyb E1	187.3	PBJ_hyb H1	204
PBJ_hyb E2	90	PBJ_hyb_H2	160.3

Day 31. 7/4/14 (Friday. ‘Murica.)

● Plan to do: Pick pHOP colonies and start liquid cultures with kan for O/N.

● Got done: Start liquid cultures

Day 32. 7/5/14 (Saturday)

● Plan to do: Freeze -> Run SEQPCR Gel on pHOP.

● Got done: pHOP gel bands are of the wrong length.

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
pHOP 1	76.4	pHOP 3	157.9
pHOP 2	128	pHOP 4	164.8

Day 33. 7/6/14 (Sunday)

● Plan to do: Get sequencing ready for PBJ_hyb_001, re run pHOP SEQPCR

● Got done: Go sequencing ready for hybrid promoter. pHOP SEQPCR gel visualization not of correct length.

Day 34. 7/7/14 (Monday)

● Plan to do: One Pot enhanced pHOP2/ Colony PCR pHOP3

● Got done: Colony PCR for pHOP3 showed wrong bands. Enhanced One Pot for pHOP2, transformed, plated on LB/Kan

Day 35. 7/8/14 (Tuesday)

● Plan to do: Start Liquid culture for pHOP2

● Got done: Started Liquid culture for pHOP2, Colony PCR pHOP2, Verified Sequencing for PBJ_hyb_001 3F, ready to go.

Day 36. 7/9/14 (Wednesday)

● Plan to do: Sequencing pcr for pHOP2, send for sequencing

● Got done: Sequencing PCR failed, do a modified digest of BBa_M30109 using BSG1, failed. Work on degradation tag design (finished)

Day 37. 7/10/14 (Thursday)

● Plan to do: Get a protocol written for testing of PBJ_hyb_001 (light and dark)

● Got done: Protocol for testing written out. Started liquid cultures in light/dark

Day 38. 7/11/14 (Friday)

● Plan to do:

● Got done: Tested hybrid and PBJ003 without chromophore. designed primers for PBJ004 (with Ptet promoter Ptet-pp+ plasmid from Dr. Moon to control EYFP).

Name	Primer
PBJ003_F	atcagtgatagagatactgagcacatctcCGGATAACAATTTCACACATACTAGAG
PBJ003_R	agggatgtcaatctctatcactgatagggaGACGAAAGGTGATACCGCTC
SEQ004_F	GTGCCAAGGGAAAAACTAGTC
SEQ004_R	gaacttcagggtcagcttg

Day 39. 7/14/14 (Monday)

● Plan to do: Begin cloning for degradation tags on PBJ003 AND PBJ_hyb_001

● Got done: Started PCR for Degradation Tags as well as Ptet-pp (same as: Ptet-T*2) minus the TetR (as a control). Designed light-box.

Day 40. 7/15/14 (Tuesday)

● Plan to do: meet with Andrew to go over light-box ordering. Continue cloning projects.

● Got done: cloning project up to Purification after Dpn1- no DNA present so re-did from PCR. Design primers for pChrom.

Name	Primers
pChrom_seq_for	GTGCCACCTGACGTCTAAG
pChrom_seq_rev	GCTGACTTGACGGGACTC

Day 41. 7/16/14 (Wednesday)

● Plan to do: Keep cloning

● Got done: kept working on pet(-)tetR, PBJ003_DegTetR, and PB_hyb_001 DegTetR up to transformation and plating

Day 42. 7/17/14 (Thursday)

● Plan to do: Check on and wrap plates. Start liquid cultures for both degradation tags, ptet(-)tetR and biobrick part from shipped frozen stock.

● Got done: Checked plates, wrapped them around 10:20 AM. Started Liquid culture at 6pm.

Day 43. 7/18/14 (Friday)

● Plan to do: Miniprep, sequencing PCR degradation tags, ptet(-). Start cloning for pChrom: Miniprep Biobrick part, digestion & ligation alongside PFM109 & transform

● Got done: Sequencing PCR for degradation tags. ptet(-) doesnt look right. Only miniprepped biobrick part.

Piece	Concentration (ng/µl)	Piece2	Concentration (ng/µl)3
Hyb degradation 1	82.3	hyb deg 2	67.1
hyb deg 3	143.4	hyb deg 4	48
PBJ 003 1	153	PBJ 003 2	61.1
PBJ 003 3	612.5	PBJ 003 4	75.1
ptet (-) tetR 1	248.8	ptet (-) tetR 2	386.4
ptet (-) tetR 3	67.6	ptet (-) tetR 4	37.7
Bba_k1017726	86.2	pFM109	8.9

Day 44. 7/19/14 (Saturday)

● Plan to do: Sequencing grad PCR for ptet(-), Sequential Digestion for pFM109, Bba_K1017726 -> Gel Extract -> Ligation -> Transform & plate

● Got done: Digested with EcoR1, Digested with SpeI, Ligation; Sequencing PCR for ptet(-): piece size not as expected ran colony pcr- band length still indicates tetR present

Day 45. 7/20/14 (Sunday)

● Plan to do: Prep DegTag products PBJ003 and hybrid for Sequencing. Transform and Plate pChrom.

● Got done: Degradation Tag on TetR for both PBJ003 and hybrid are ready for sequencing. Transform and plate pChrom on lb/kanR plates. Put in 37 at 7:50pm

Day 46. 7/21/14 (Monday)

● Plan to do: send tags off for sequencing. Start ptet(-) tetR cloning steps. Start liquid cultures for Chromophore project.

● Got done: tags sent for sequencing. finished from PCR to dpn1 on ptet (-) tetR. colony pcr for chromophore failed. Redo ligation and transformation

Day 47. 7/22/14 (Tuesday)

● Plan to do: redo colony pcr for chromophore. Work on ptet(-)tetR positive control.

● Got done: kept working on ptet(-)tetR positive control- dpn1 and purify,transformed and plated. Not enough colonies for chromophore to colony pcr. Started liquid culture at night.

Day 48. 7/23/14 (Wednesday)

● Plan to do: Miniprep and sequencing pcr for chromophore. Check and wrap plates, start liquid cultures around 8 or 9.

● Got done: Miniprep and Sequencing PCR for chromophore. Prepped for sequencingstarted (-)tetR liquid cultures. worked on PCR seq for Degradation tags on PBJ003 and Phybrid. “need” to purify before sending pcr product.

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
pChrom #1	142.2	pChrom #2	122.8
pChrom #3	112.8	pChrom #4	78.7

Day 49. 7/24/14 (Thursday)

● Plan to do: Purify PCR products. Miniprep and check sequencing pcr for (-)tetR. Send all for sequencing

● Got done: Purified PCR product for chrom and degradation tags. Prepped for sequencing. Miniprep and run sequencing PCR for (-)tetR (no bands)

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
(-) tetR 1	102	(-) tetR 2	129.6
(-) tetR 3	455.3	(-) tetR 4	906.4

Day 50. 7/25/14 (Friday)

● Plan to do: Gradient SEQPCR on (-)tetR, run a gel visualizatoin, prep for sequencing.

● Got done: Ran gradient on (-)tetR #3 and 4 with gel visualization, saw bands at wrong length. doing colony PCR tomorrow.

Day 51. 7/26/14 (Saturday)

● Plan to do: Brute Force Colony PCR (basically picking 32 colonies!). Freeze and Miniprep correct ones.

● Got done: Brute Force Colony PCR and gel visualization finished.

Day 52. 7/27/14 (Sunday)

● Plan to do: Start all cloning procedures by taking #4 of the degradation tags from hybrid and pBJ003. Prep for sequencing the miniprepped (-)tetR DNA samples (#9,12,15,16).

● Got done: Prep (-)tetR for sequencing. Start cloning PBJ004 and PBJ_hyb_002

Day 53. 7/28/14 (Monday)

● Plan to do: Digest Biobrick part to check for correct lengths… continue cloning of PBJ004 and PBJ_hyb_002 with the new ptet promoter insertion. Look more at LED grid coding.

● Got done: Digestion of biobrick part only showed a smear. Still send both miniprep and pcr product for sequencing

Day 54. 7/29/14 (Tuesday)

● Plan to do: Check plates in the morning, wrap. Start cloning for PBJ005 and Hybrid 003 which are PBJ003 and Hybrid001 plasmids that just have the new ptet promoter inserted. Start liquid cultures at night.

● Got done: plates checked and wrapped. Sanity check #2 for biobrick part, doing a double digestion. Started cloning of PBJ005 and Hybrid003. Liquid cultures started. Retransform003, order new sequencing primer, redo double digestion.

Day 55. 7/30/14 (Wednesday)

● Plan to do: Freeze, miniprep, send off for sequencing of PBJ004 and Hybrid002. Delete the CAC in the maps for PBJ003 with DegTagTetR and PBJ_hyb with DegTagTetR. Get iGEM calendar set up. Email Pakrasi and Moon about setting up final iGEM meeting

● Got done: Start Modeling of light promoter system. (-) tetR still no luck. Sequence verified pCHROM #1. Freeze miniprep, PBJ004, Hybrid 002. Start liquid cultures for PBJ005, Hybrid 003.

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
PBJ004 #1	145.6	PBJ004 #2	165.4
PBJ004 #3	97.7	PBJ004 #4	122.3
Hybrid002 #1	80	Hybrid002 #2	436.8
Hybrid002 #3	102.4	Hybrid002 #4	117.5

Day 56. 7/31/14 (Thursday)

● Plan to do: Make LB/ (spec + cm) & (spec + kan) plates. Seq PCR for (-) tetR. Freeze miniprep PBJ005, Hybrid003. Send PBJ004, PBJ005, Hybrid 003, Hybrid 002, (-) tetR for sequencing. Design experimental protocol. Transform.

● Got done: Made plates. Miniprep PBJ005, hybrid 003. Sequencing PCR, gel visualization for PBJ005, hybrid003, (-) tetR. Gel visualize. Gradient PCR hybrid003, (-)tetR, visualize. Found Hybrid003 1E, 1F, (-)tetR wrong bands. Prep PBJ004, PBJ005, hybrid002 for sequencing. Transform/ plate pchrom + (pos/neg/pbj003,phybrid).

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
PBJ005 #1	145.6	PBJ005 #2	165.4
PBJ005 #3	97.7	PBJ005 #4	122.3
Hybrid003 #1	80	Hybrid003 #2	436.8
Hybrid003 #3	102.4	Hybrid003 #4	117.5

Day 57. 8/1/14 (Friday)

● Plan to do: Prep Hybrid003 for sequencing. Send all in before 9. Pick cotransformed colonies. Start liquid culture.

● Got done: Sent for sequencing. Colonies not growing well. Colony PCR pchrom3 (from 7/29/14). Work on modeling.

Day 58. 8/2/14 (Saturday)

● Plan to do: Check plates. Wrap. Start liquid cultures

● Got done: Wrap colony PCR plate. Transformants not growing. *** Realization, pCHROM has kan resistance, so using the wrong plates the whole time. Started liquid cultures for pCHROM3, pCHROM1.

Day 59. 8/3/14 (Sunday)

● Plan to do: Miniprep pCHROM3 and 1

● Got done: Freeze pCHROM3, Miniprep pCHROM3 and 1. Sequencing results for new Ptet bad

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
pChrom 1	157.5	pChrom3 #4	133.6
pChrom3 #5	102.6	pChrom3 #12	79.4
pChrom3 #13	91.5

Day 60. 8/4/14 (Monday)

● Plan to do: Make Kan/CM plates. Check Sequence for pCHROM3. Transform and plate for experiment

● Got done: Made plates. Sequence PCR #1 showed multiple bands. Sequence PCR #2 showed wrong bands. Started new Ptet stuff over again from PCR. Transformed and plate experimental stuff

Day 61. 8/5/14 (Tuesday)

● Plan to do: Wrap Plates. Miniprep PBJ003, PBJ_Hyb_001. Redo cloning for new Ptet plasmids. Start liquid cultures for experimentation. Work on presentation

● Got done: Wrap plates. Miniprep. Redo PBJ005

Day 62. 8/6/14 (Wednesday)

● Plan to do: Run experiment. Induce. Check data

● Got done: Lighted growth chamber in use at 30 degrees. Cheryl advise to abort util tomorrow… Work on presentation. Purify, Transform PBJ005 redo. Design PBJpos

Day 63. 8/7/14 (Thursday)

● Plan to do: Run experiment. Induce. Check data

● Got done: Ran experiment. Dilute to OD .1, grow for 2 hours, induction. Start PBJ005 liquid cultures, measure fluorescence. NO FLUORESCENCE. Plan out new positive control, knocking out CcaR/CcaS/Pcpcg2/tetR from PBJ003.

Name	Primer
PBJpos_F	GAGCGGTATCACCTTTCGTC
PBJpos_R	CCTCGAGCTTAGGACGTCG
SEQpos_F	GTTCTTTACGATGCCATTGG
SEQpos_R	CACCATCTAGTATTTCTCCTCTTTC

Day 64. 8/8/14 (Friday)

● Plan to do: Send PBJ005 for sequencing

● Got done: Froze, miniprep, Sequencing PCR. Send Purified Seq PCR product for sequencing. Jeffrey’s last day

Day 65. 8/11/14 (Monday)

● Plan to do: Start cloning for PBJpos

● Got done: PCR, visualize, DPN1, purify, Blunt end ligate. Transform PBJpos. Also Transform PSL2264

Day 66. 8/12/14 (Tuesday)

● Plan to do: Start liquid culture for PBJpos, PSL2264

● Got done: Wrapped plates. Checked sequence results for PBJ005=bad. Started liquid cultures for PBJpos

Day 67. 8/13/14 (Wednesday)

● Plan to do: Miniprep, send for sequencing, start liquid cultures for experimentation

● Got done: Miniprepped and sent PBJpos #1-#4 for sequencing. Started liquid cultures for pSL2264, PBJpos 1-4, current positive control and negative control for experimentation tomorrow.

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
PBJpos #1	360.5	PBJpos #2	349.9
PBJpos #3	443.9	PBJpos #4	270.8

Day 68. 8/14/14 (Thursday)

● Plan to do: Run fluorescence tests for old and new positive controls. Sanity check for pSL2264

● Got done: pSL2264 not fluorescing as expected. PBJpos slightly better, Old positive control and negative control as expected. Induce pSL2264 with IPTG. Design new PBJ003 with deletion of competing TSS

Day 69. 8/15/14 (Friday)

● Plan to do: Design primers for new PBJ plasmids with modified RBS. Plan out fall semester work schedule.

● Got done: Design primers for new PBJ plasmids with modified RBS.

Day 70. 8/25/14 (Monday)

● Plan to do: Start cloning for PBJposrbs (new positive control with enhanced RBS) until transformation

● Got done: PCR, gel visualize, DPN1, Ligation, Transformation, Plating. Work on Protocol section for website.

Day 71. 8/26/14 (Tuesday)

● Plan to do: Start Liquid Culture

● Got done: Start liquid culture. Work on protocols.

Day 72. 8/27/14 (Wednesday)

● Plan to do: Miniprep. Run sequencing PCR, send for sequencing

● Got done: Miniprep. Run sequencing PCR, results bad. Rerun sequencing PCR with correct primers.

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
PBJpos_rbs #1	166.3	PBJpos_rbs #2	165.2
PBJpos_rbs #3	107.9	PBJpos_rbs #4	271.5

Day 73. 8/28/14 (Thursday)

● Plan to do: Visualize sequencing PCR. PCR purify, prep for sequencing. Send

● Got done: visualize sequencing PCR. PCR purify, prep for sequencing. Work on title and abstract

Day 74. 8/29/14 (Friday)

● Plan to do: Send in for sequencing (morning). Make electrocompetent cells

● Got done: Send in for sequencing. Make electrocompetent cells. Work on title and abstract.

Day 75. 9/1/14 (Monday)

● Plan to do: Sequence verify PBJpos_RBS. Start liquid cultures for experimentation.

● Got done: Sequence verify PBJpos_RBS. Start liquid cultures for experimentation. Final safety forms due.!

Day 76. 9/2/14 (Tuesday)

● Plan to do: Test for Fluorescence of newest positive control.

● Got done: tested for fluorescence. Dilute to 0.1 grew an extra 6 hours, tested again. EYFP did not fluoresce as expected. Compard 5’ UTRs between different positive controls. redesign primers in order to change pTet from tabor to moon’s.

Name	Primer
PBJptet_F	tagagatactgagcacatctcCGGATAACAATTTCACACATACTAGAG
Pbjptet_R	tcactgatagggatgtcaatctctatcactgatagggaACGAAAGGTGATACCGCTCG
PBJpos_seq_F	Use PBJRBS seq_F
PBJpos_seq_R	Use PBJRBS seq_R
BbaMoonptet_F	Use PBJptet_F
BBaMoonptet_R	tcactgatagggatgtcaatctctatcactgatagggaAACTGCTTTCGGTTAGACTTC
BbaMoon_seq_f	AACCGGTAACAGTACAAATCC
BbaMoon_seq_r	Use PBJRBS seq_R

Day 77. 9/4/14 (Thursday)

● Plan to do: Start cloning once primers are received.

● Got done: Organize primers for assembly. Start cloning for swapping Ptet on positive controls.

Day 78. 9/5/14 (Friday)

● Plan to do: Take cloning through transformation

● Got done: Gel, Digestion, Purification, Ligation, Transformation

Day 79. 9/6/14 (Saturday)

● Plan to do: Start liquid cultues

● Got done: Wrapped plates in the morning. Start liquid cultures at night

Day 80. 9/7/14 (Sunday)

● Plan to do: Miniprep, Sequencing PCR, Prep for sequencing.

● Got done: Miniprep, Sequencing PCR, Prepare for sequencing and send off tomorrow morning.

Piece	Concentration (ng/µl)	Piece	Concentration (ng/µl)
PBJposmoon #1	82.9	PBJposmoon #2	179.4
PBJposmoon #3	91.1	PBJposmoon #4	50.5
Bbaposmoon #1	84.9	Bbaposmoon #2	180.3
Bbaposmoon #3	72.7	Bbaposmoon #4	28.5

Day 81. 9/10/14 (Wednesday)

● Plan to do: Check on sequences

● Got done: Verified sequence for Bba_pos_moon. Started liquid cultures for experimentation tomorrow. Start designing primers for biobricking.

Day 82. 9/11/14 (Thursday)

● Plan to do: Check for fluorescence of new pTet. Finish and clarify design of primers for biobricking.

● Got done: Bba_pos_moon fluoresces as expected!!!!! Designed primers to start biobricking. Sent for ordering.

Name	Primers
Bba_PBJ_f	gaattcgcggccgcttctagagAGCCCATTGTGCTTTTC
Bba_PBJ_r	ctgcagcggccgctactagtaGATTATTAGGACCCACTTTCAC
PBJ_seq_prefix_f	CCTATAAAAATAGGCGTATCAC
PBJ_seq_prefix_r	GCAAAAGTGAGTATGGTG
PBJ_seq_Suffix_f	CTTTGAGTGAGCTGATACC
PBJ_seq_Suffix_r	GATTAACAGCGCATTAGAG
Bba_pos_f	gaattcgcggccgcttctagagGAAGTCTAACCGAAAGCAG
Bba_pos_r	actagtaAGTAGAGAGCGTTCACCG
Bba_seq_prefix_f	Use PBJ_seq_prefix_f
Bba_seq_prefix_r	TGTTCTGCTGGTAGTGGTC
Bba_seq_suffix_f	Use PBJ_seq_suffix_f
Bba_seq_suffix_r	CACAAGCTGGAGTACAACTAC

Team:WashU StLouis/Notebook/Brauer

From 2014.igem.org

Latest revision as of 15:49, 15 September 2014

Brauer Log

Brauer Notebook

Special thanks to our 2014 Sponsors:

If you want to sponsor us, please contact us below!

@@ Line 1: / Line 1: @@
-{{:Team:WashU_StLouis/CSS}}
 {{WashU/Layout}}
+{{:Team:WashU_StLouis/CSS}}
 <html>
 <style>
 #contentSub, #footer-box, #catlinks, #search-controls, #p-logo, .printfooter, .firstHeading,.visualClear {display: none;} /*-- hides default wiki settings --*/
+div.floating-menu {position:fixed;background:#fff4c8;border:1px solid #ffcc00;width:150px;z-index:100;}
+div.floating-menu a, div.floating-menu h3 {display:block;margin:0 0.5em;}
 </style>
+<div class="floating-menu">
+     <h3>Brauer Log</h3>
+     <a href="#1">Week 1</a>
+     <a href="#2">Week 2</a>
+     <a href="#3">Week 3</a>
+     <a href="#4">Week 4</a>
+     <a href="#5">Week 5</a>
+     <a href="#6">Week 6</a>
+     <a href="#7">Week 7</a>
+     <a href="#8">Week 8</a>
+     <a href="#9">Week 9</a>
+     <a href="#10">Week 10</a>
+     <a href="#11">Week 11</a>
+     <a href="#12">Week 12</a>
+     <a href="#13">Week 13</a>
+     <a href="#14">Week 14</a>
+</div>
 <!--  here ends the section that changes the default wiki template to a white full width background -->
@@ Line 17: / Line 37: @@
 <h1> <center><b> Brauer Notebook </b> </center></h1>
+<body bgcolor=white lang=EN-US style='tab-interval:.5in'>
+<div class=WordSection1>
+<p class=MsoNormal align=center style='text-align:center;line-height:normal'><span
+class=GramE><b style='mso-bidi-font-weight:normal'>Captains’ Log.</b></span><b
+style='mso-bidi-font-weight:normal'> <span class=GramE>iGEM</span> 2014. Team <span
+class=SpellE>NitroGENIUS</span>. The <span class=SpellE>Brauer</span> Group.</b></p>
+<p class=MsoNormal align=center style='text-align:center;line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal align=center style='text-align:center;line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="1"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 1. 6/2/2014 (Monday) </b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l11 level1 lfo1'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> design
+primers/plasmids </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l11 level1 lfo1'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: started
+primer/plasmid design</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 2. 6/3/2014 (Tuesday) </b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l33 level1 lfo2'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Create and
+work on presentation</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l33 level1 lfo2'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Monsanto
+Presentation</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 3. 6/4/2014 (Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l27 level1 lfo3'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Monsanto
+Visit</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l27 level1 lfo3'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Monsanto
+Visit</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 4. 6/5/014 (Thursday) </b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l5 level1 lfo4'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> More plasmid
+editing/primer design. Edit website. Plan out video series.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l5 level1 lfo4'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Finished <span
+class=SpellE>pLight</span> plasmid design and primers, sent to Cheryl for
+verification. Typed up brainstorming for video sessions, contacted WUTV about
+using equipment, contacted <span class=SpellE>WashU</span> Assembly Series
+director for permission to allow a Monsanto scientist to give a lecture during
+the Fall 2014 semester</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 5. 6/6/2014 (Friday) </b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l3 level1 lfo5'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Get iGEM
+safety forms filled out. Go over primer/plasmid design. Order primers. Get
+keys/access to <span class=SpellE>Brauer</span> Hall. Design a timeline for the
+summer. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l3 level1 lfo5'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Finished design
+of plasmid PBJ001. Finished all (including sequencing) primer design. Take
+another look on Sunday, 6/8/2014.</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=419
+ style='margin-left:4.65pt;border-collapse:collapse;mso-table-layout-alt:fixed;
+ mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:15.0pt'>
+  <td width=106 nowrap valign=bottom style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Name<o:p></o:p></span></b></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Primer<o:p></o:p></span></b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>EYFP-&gt; ColE1 _f<o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ <span class=SpellE>tttttttgctcttcaaatGGGATAACGCAGGAAAGAAC</span>
+’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>EYFP-&gt; ColE1 _r<o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ <span class=SpellE>tttttttgctcttcagtgCTGAAGGCTGACAAAATCTATACTG</span>
+’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:15.0pt'>
+  <td width=106 nowrap valign=bottom style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Pcpcg2 -&gt; <span class=SpellE>Ptet_f</span><o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ <span class=SpellE>tttttttgctcttcaTTCCCAGTCACGACGTTG</span>
+’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Pcpcg2 -&gt; <span class=SpellE>Ptet_r</span><o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ <span class=SpellE>tttttttgctcttcaTCGGTCAGTGCGTCCTG</span>
+’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>TetR_f</span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’tttttttgctcttcaCACCTACAACAAAGCTCTCATC 3’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>TetR_r</span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’tttttttgctcttcagaaGGCTAGCTCAGTCCTAGGTACAG 3’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>EYFP_f</span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ <span class=SpellE>tttttttgctcttcaattCTTGTGCAATGTAACATCAGAG</span>
+’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:8;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>EYFP_r</span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ <span class=SpellE>tttttttgctcttcacgaGTGGAATTGTGAGCGGATAAC</span>
+’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:9;height:15.0pt'>
+  <td width=106 nowrap valign=bottom style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Seq1_f<o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ ACAAATCCGCCGCCCTAG 3’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:10;height:15.0pt'>
+  <td width=106 nowrap valign=bottom style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Seq1_r<o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ GACTTGAGCGTCGATTTTTGTG 3’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:11;height:15.0pt'>
+  <td width=106 nowrap valign=bottom style='width:106.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Seq2_f<o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ GGCTTTTTTACCGGCTAGTTTC 3’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:12;mso-yfti-lastrow:yes;height:15.0pt'>
+  <td width=106 nowrap valign=bottom style='width:106.0pt;border:solid black 1.0pt;
+  border-right:none;mso-border-top-alt:solid black .5pt;mso-border-left-alt:
+  solid black .5pt;mso-border-bottom-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Seq2_r<o:p></o:p></span></p>
+  </td>
+  <td width=313 nowrap valign=bottom style='width:313.25pt;border:solid black 1.0pt;
+  border-left:none;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ GATTTTCTCCCCCTTTTCTTC 3’<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="2"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 6. 6/9/2014 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l0 level1 lfo6'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Get iGEM
+safety forms filled out. Order primers. Get keys/access to <span class=SpellE>Brauer</span>
+Hall. Design a timeline for the summer. Complete lab safety forms for <span
+class=SpellE>WashU</span>. Design primers/plasmid map for positive/negative
+control. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l0 level1 lfo6'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Ordered
+primers (should arrive by Wednesday at the latest). Got keys/access to <span
+class=SpellE>Brauer</span> Hall. Completed lab safety forms for <span
+class=SpellE>WashU</span>. </p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 7. 6/10/2014 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l12 level1 lfo7'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Finish
+design of <span class=SpellE>pos</span>/<span class=SpellE>neg</span> control
+plasmids. Print <span class=SpellE>Brauer</span> lab safety form. Finalize
+proposal and send to Penn State iGEM contact along with project presentation.
+Design light experiment. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l12 level1 lfo7'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Designed
+primers for the positive and negative control plasmids. Printed safety form.
+Sent Penn State iGEM project info. PCR, ran visualization, and extracted
+pieces.</p>
+<div align=center>
+<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0 width=489
+ style='margin-left:4.65pt;border-collapse:collapse;mso-table-layout-alt:fixed;
+ border:none;mso-border-alt:solid black .5pt;mso-yfti-tbllook:1184;mso-padding-alt:
+in 5.4pt 0in 5.4pt;mso-border-insideh:.5pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border:solid black 1.0pt;
+  border-right:none;mso-border-top-alt:solid black .5pt;mso-border-left-alt:
+  solid black .5pt;mso-border-bottom-alt:solid black .5pt;background:black;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Name<o:p></o:p></span></b></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;background:black;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Primer<o:p></o:p></span></b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>PJT122_modNeg_minuslacZ1_Forward<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GGTTTTCCCAGTCACGACG<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>PJT122_modNeg_minuslacZ1_Reverse<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>CTGGAGCCCGTCAGTATCG <o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>PJT122_modNeg_minuscph8_2_Forward<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GCAGGGCATGACAAAAGAAG </span><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>PJT122_modNeg_minuscph8_2_Reverse<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>CTGAGTTGTACGGTGGTGGC <o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>PJT122_modNeg_seq_cph8_Foward<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>CAACGTGCCAACCAGTTTTAG <o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>PJT122_modNeg_seq_cph8_Reverse<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GAACGACCTACACCGAACTG </span><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>PJT122_modNeg_seq_lacZ_Forward<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>CAAAAACGCCGTCAATTTG <o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:8;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>PJT122_modNeg_seq_lacZ_Reverse<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GCTTTACACCGTTGATATTGC <o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:9;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>pSL2264</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>_EYFP_pos_Foward<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>tttttttgctcttcaCGTGGCTTTGTTGAATAAATC</span></span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:10;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>pSL2264</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>_EYFP_pos_Reverse<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>tttttttgctcttcatcaAGCGGATAACAATTTCACACATAC</span></span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:11;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>pTetR100</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>_pos_Forward<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>tttttttgctcttcaTGAGCTAGCACTGTACCTAGGAC</span></span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'> <o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:12;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>pTetR100</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>_pos_Reverse<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>tttttttgctcttcaacgCTGAAGTGGCGGAACTGC</span></span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'> </span><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:13;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>pTetEYFP03</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>_seq_Forward<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GGTCATGGTAAAGGCAATGG <o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:14;mso-yfti-lastrow:yes;height:15.0pt'>
+  <td width=211 nowrap valign=bottom style='width:210.75pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>pTetEYFP03</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>_seq_Reverse<o:p></o:p></span></p>
+  </td>
+  <td width=278 nowrap valign=bottom style='width:278.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>TATGGCAATGGCACAATTAGATG <o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 8. 6/11/2014 (Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l35 level1 lfo8'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Moon Lab
+Group meeting at 3PM. Work on finishing Gel DNA extraction, Dpn1, Purification,
+then One-Pot, then finally Transformation and plating (2 LB/cm plates). [<b
+style='mso-bidi-font-weight:normal'>Realization! </b>We discovered today that
+in order for the <span class=SpellE>ccaS</span>/<span class=SpellE>ccaR</span>
+light system to function correctly, we need the PCC6803 genes <i
+style='mso-bidi-font-style:normal'>ho1 </i>and <span class=SpellE><span
+class=GramE><i style='mso-bidi-font-style:normal'>pcyA</i></span></span><span
+class=GramE> which</span> are needed to <span class=SpellE>heme</span> into PCB
+(a chromophore) via a two step oxidation/reduction process. Ray is working on
+this aspect and we are planning to help/take over this part of his work. Our
+plan is to still transform plasmid PBJ and send it off for sequencing to make
+sure we have our designed plasmid. The next step would be to co-transform the
+two plasmids, one containing the <i style='mso-bidi-font-style:normal'>ho1 </i>and
+<span class=SpellE><i style='mso-bidi-font-style:normal'>pcyA</i></span> genes
+and the plasmid PBJ into <i style='mso-bidi-font-style:normal'>E. coli</i> and
+after sending off for sequencing, test the light promoter system. ]</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l35 level1 lfo8'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Finished
+Gel DNA extraction and measured concentration using <span class=SpellE>nanoquant</span>
+<span class=SpellE>tecan</span>. Did not need to do Dpn1 nor purification. Put
+in one-pot after 12 <span class=GramE>noon</span>. Transformed and incubated
+O/N. PCR for Ray’s <span class=SpellE>Biobrick</span> plasmid for <i
+style='mso-bidi-font-style:normal'>ho1 </i>and <span class=SpellE><i
+style='mso-bidi-font-style:normal'>pcyA</i></span> parts. </p>
+<div align=center>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='margin-left:.5in;border-collapse:collapse;border:none;mso-border-alt:
+ solid windowtext .5pt;mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=108 valign=top style='width:107.75pt;border:solid windowtext 1.0pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Piece</p>
+  </td>
+  <td width=123 valign=top style='width:122.65pt;border:solid windowtext 1.0pt;
+  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Concentration (<span
+  class=SpellE>ng</span>/µl)</p>
+  </td>
+  <td width=86 valign=top style='width:85.5pt;border:solid windowtext 1.0pt;
+  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Piece</p>
+  </td>
+  <td width=127 valign=top style='width:126.9pt;border:solid windowtext 1.0pt;
+  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Concentration (<span
+  class=SpellE>ng</span>/µl)</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=108 valign=top style='width:107.75pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>TetR</span></p>
+  </td>
+  <td width=123 valign=top style='width:122.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>143.1</p>
+  </td>
+  <td width=86 valign=top style='width:85.5pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>EYFP</p>
+  </td>
+  <td width=127 valign=top style='width:126.9pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>128.4</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=108 valign=top style='width:107.75pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Col A</p>
+  </td>
+  <td width=123 valign=top style='width:122.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>12.3</p>
+  </td>
+  <td width=86 valign=top style='width:85.5pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Col B</p>
+  </td>
+  <td width=127 valign=top style='width:126.9pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>5.8</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=108 valign=top style='width:107.75pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Col D</p>
+  </td>
+  <td width=123 valign=top style='width:122.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>12.9</p>
+  </td>
+  <td width=86 valign=top style='width:85.5pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Col E</p>
+  </td>
+  <td width=127 valign=top style='width:126.9pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>2.8</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4'>
+  <td width=108 valign=top style='width:107.75pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>G2 A</p>
+  </td>
+  <td width=123 valign=top style='width:122.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>26.2</p>
+  </td>
+  <td width=86 valign=top style='width:85.5pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>G2 B</p>
+  </td>
+  <td width=127 valign=top style='width:126.9pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>26.2</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5'>
+  <td width=108 valign=top style='width:107.75pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>G2 C</p>
+  </td>
+  <td width=123 valign=top style='width:122.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>45.4</p>
+  </td>
+  <td width=86 valign=top style='width:85.5pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>G2 D</p>
+  </td>
+  <td width=127 valign=top style='width:126.9pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>26.6</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6;mso-yfti-lastrow:yes'>
+  <td width=108 valign=top style='width:107.75pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>G2 E</p>
+  </td>
+  <td width=123 valign=top style='width:122.65pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>15.3</p>
+  </td>
+  <td width=86 valign=top style='width:85.5pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>G2 F</p>
+  </td>
+  <td width=127 valign=top style='width:126.9pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>77.7</p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 9. 6/12/2014 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l35 level1 lfo8'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Gel extract,
+one-pot for Ray’s plasmid then transform and plate. <span class=GramE>pick</span>
+PBJ002 colonies, start O/N liquid cultures. Work on negative and pos. control
+once primers arrive (check around noon). Also, grow a culture of Ray’s <span
+class=SpellE>BbaP</span> DNA (<span class=SpellE>AmpR</span>) as well as
+Cheryl’s <span class=SpellE>pTet</span> 100-T (where we get the <span
+class=SpellE>TetR</span> from)</p>
+<p class=MsoNormal style='line-height:normal'><span style='mso-tab-count:1'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>Plan
+out <span class=SpellE>seq</span> PCR for PBJ002.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l35 level1 lfo8'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Gel
+extract and one pot for Ray’s plasmid. Re-ran PCR for Rays <span class=SpellE>BbaP</span>
+(<span class=SpellE>SpeC_AraC_Bba</span>) plasmid. Gel extract on new <span
+class=SpellE>BbaP</span> part. Did not yield bands- only saw a <span
+class=GramE>smear.</span> Received primers for <span class=SpellE>neg</span>
+and <span class=SpellE>pos</span> controls- mixed appropriate primer
+concentrations. Cheryl started <span class=SpellE>pTet</span> liquid culture.
+Started <span class=SpellE>BbaP</span> liquid cultures. <span class=SpellE>Electroporated</span>
+original <span class=SpellE>Bbap</span>. Plated on 2 <span class=SpellE>specR</span>
+plates (100 <span class=SpellE>uL</span>) and (the rest). <span class=GramE>put</span>
+into incubator. </p>
+<p class=MsoNormal style='line-height:normal'><span style='mso-tab-count:1'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>Planned
+out <span class=SpellE>seq</span> PCR for PBJ002. </p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 10. 6/3/2014 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l35 level1 lfo8'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> meet at 9:15
+in the ICARES to set up <span class=GramE>video conference</span> with the Penn
+State. </p>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'>1<span
+style='color:#38761D'>. PBJ002: <span class=SpellE>Miniprep</span> -&gt; PCR
+with sequencing primers before ICARES meeting</span></p>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='margin-left:1.0in;line-height:normal'><span
+class=SpellE><span class=GramE><span style='color:#38761D'>miniprep</span></span></span><span
+style='color:#38761D'> <span class=SpellE>pTet</span> and <span class=SpellE>BbaP</span>
+we made liquid cultures for yesterday. <span class=SpellE><span class=GramE>miniprep</span></span>
+this along with the PBJ <span class=SpellE>miniprep</span>. </span></p>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'>2. <span
+class=GramE><span style='color:#38761D'>pour</span></span><span
+style='color:#38761D'> gel for visual confirmation </span></p>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'><b
+style='mso-bidi-font-weight:normal'><span style='color:#38761D'>...ICARES
+meeting… then...</span></b></p>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'>3. <span
+class=GramE><span style='color:#38761D'>take</span></span><span
+style='color:#38761D'> PCR product and run a gel (10 <span class=SpellE>uL</span>
+DNA, 2uL 6X dye) to check. 30 minutes.</span></p>
+<p class=MsoNormal style='margin-left:1.0in;line-height:normal'><span
+style='color:#38761D'>Observe which PCR product in gel (which lane from tube)
+is of the right length.</span></p>
+<p class=MsoNormal style='margin-left:1.0in;line-height:normal'><s>Purify PCR
+product.</s></p>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'><b
+style='mso-bidi-font-weight:normal'><span style='color:#38761D'>Send <span
+class=SpellE>miniprepped</span> <span class=GramE>plasmid<span
+style="mso-spacerun:yes">&nbsp; </span><s>AND</s></span><s> purified PCR</s>
+off for sequencing of PBJ002. </span></b></p>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'>4. <span
+class=GramE>work</span> on both the Positive and negative controls.</p>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'>5.<span
+style='color:#38761D'> Pick colonies and start O/N culture for <span
+class=SpellE>Spec_AraC_Bba</span>. </span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l35 level1 lfo8'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: From the
+above mentioned steps: Steps 1- 5. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l35 level2 lfo8'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>For the <span class=SpellE>Pos</span> and <span
+class=SpellE>Neg</span> controls, we did PCR -&gt; <span class=SpellE>nanoquant</span>
+measure after purification step (reference <span class=SpellE>pos</span>/<span
+class=SpellE>neg</span> control step documents) <span class=SpellE><span
+style='color:red'>TetR</span></span><span style='color:red'> backbone not
+present. Re-run tomorrow. </span></p>
+<div align=center>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
+ mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Piece</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Concentration (<span
+  class=SpellE>ng</span>/µl)</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Piece</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Concentration (<span
+  class=SpellE>ng</span>/µl)</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>PBJ 02 #1</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>103.9</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>PBJ 02 #2</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>90.1</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>PBJ 03 #3</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>137.5</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>PBJ 02 #4</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>76.9</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>Ptet</span></p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>60.4</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>BbaP</span></p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>62.2</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>(-) <span class=SpellE><span
+  class=GramE>lacZ</span></span> #2</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>103.9</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>(-) <span class=SpellE><span
+  class=GramE>lacZ</span></span> #6</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>90.1</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>(-) <span class=SpellE><span
+  class=GramE>lacZ</span></span> #3</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>137.5</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>(-) <span class=SpellE><span
+  class=GramE>lacZ</span></span> #7</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>76.9</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>(-) <span class=SpellE><span
+  class=GramE>lacZ</span></span> #5</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>60.4</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>(-) <span class=SpellE><span
+  class=GramE>lacZ</span></span> #8</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'>62.2</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7;mso-yfti-lastrow:yes;height:11.8pt;mso-row-margin-right:
+.4pt'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:11.8pt'>
+  <p class=MsoNormal style='margin-left:5.4pt;line-height:normal'>EYFP</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:11.8pt'>
+  <p class=MsoNormal align=right style='margin-left:5.4pt;text-align:right;
+  line-height:normal'>34.6</p>
+  </td>
+  <td style='mso-cell-special:placeholder;border:none;padding:0in 0in 0in 0in'
+  width=239 colspan=2><p class='MsoNormal'>&nbsp;</td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 11. 6/14/2014 (Saturday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level1 lfo9'><![if !supportLists]><span
+style='color:red'><span style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span style='color:red'>Plan to do: <o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=SpellE>Miniprep</span> -&gt; Visualize <span
+class=SpellE>seq</span> PCR product of <span class=SpellE>Spec_AraC_Bba</span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level3 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=GramE>could</span> not run <span
+class=SpellE>seq</span> PCR on this because <span class=SpellE>seq</span>
+primers do not work. Re-design sequencing primers and order them.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='color:red'><span style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span style='color:red'>Re-run PCR for <span
+class=SpellE>TetR</span> backbone. Failed yesterday. <o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='color:red'><span style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><s><span style='color:red'>One pot <span
+class=SpellE>Pos</span> control<o:p></o:p></span></s></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>One pot <span class=SpellE>Neg</span> control</p>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'>...Later...</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Transform/plate <span class=SpellE>neg</span> control</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><s>Transform/plate <span class=SpellE>pos</span>
+control<o:p></o:p></s></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level1 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=SpellE>Miniprepped</span> and measured
+concentration on SAB product. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level3 lfo9'><![if !supportLists]><span
+style='background:yellow;mso-highlight:yellow'><span style='mso-list:Ignore'>&#9632;<span
+style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+style='background:yellow;mso-highlight:yellow'>Project SAB is on hold until
+primers arrive.<o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Re ran PCR on the <span class=SpellE>TetR</span>
+backbone. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=GramE>sent</span> new redesigned sequencing
+primers for SAB to Ray for ordering.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Gel Extraction/Recovery on <span class=SpellE>TetR</span>
+backbone PCR product </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Dpn1 for <span class=SpellE>TetR</span> backbone </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=SpellE>TetR</span> purification and
+measurement= 4.4 <span class=SpellE>ng</span>/<span class=SpellE>uL</span> </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level3 lfo9'><![if !supportLists]><span
+style='background:yellow;mso-highlight:yellow'><span style='mso-list:Ignore'>&#9632;<span
+style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+style='background:yellow;mso-highlight:yellow'>Project Positive Control is on
+hold until we can re-evaluate the primers and find the error/form a new plan of
+approach.<o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level3 lfo9'><![if !supportLists]><span
+style='color:blue'><span style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span style='color:blue'>Correction: re-did PCR
+for <span class=SpellE>TetR</span> backbone again. (<span class=GramE>was</span>
+the reason we couldn’t get it to work before because we used the wrong
+plasmid?)<o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:2.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level4 lfo9'><![if !supportLists]><span
+style='color:blue'><span style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span class=GramE><span style='color:blue'>gel</span></span><span
+style='color:blue'> only yielded bands indicating primers.<o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='color:#38761D'><span style='mso-list:Ignore'>&#9675;<span
+style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+style='color:#38761D'>One pot Negative control (-) <span class=SpellE>lacZ</span><o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l39 level2 lfo9'><![if !supportLists]><span
+style='color:#38761D'><span style='mso-list:Ignore'>&#9675;<span
+style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+style='color:#38761D'>Transform/plated Negative control (-) <span class=SpellE>lacZ</span><o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+line-height:normal'><span style='color:#38761D'><o:p>&nbsp;</o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+line-height:normal'><span style='color:#38761D'><o:p>&nbsp;</o:p></span></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 12. 6/15/2014 (Sunday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l8 level1 lfo10'><![if !supportLists]><span
+style='color:red'><span style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span style='color:red'>Plan to do: <o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l8 level2 lfo10'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=SpellE>TetR</span> backbone for positive
+control run from PCR- &gt;Transformation/Plating</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l8 level2 lfo10'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Pick <span class=SpellE>Neg</span> Control colonies. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l8 level3 lfo10'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Start liquid cultures for O/N </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l8 level1 lfo10'><![if !supportLists]><span
+style='color:#38761D'><span style='mso-list:Ignore'>&#9679;<span
+style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+style='color:#38761D'>Got done</span>:<span style='color:#38761D'><o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l8 level2 lfo10'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>(+) <span class=SpellE>TetR</span> backbone from
+TetR-100T <span class=SpellE>miniprepped</span> plasmid: PCR -&gt; DNA
+purification</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l8 level2 lfo10'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Picked <span class=SpellE>Neg</span> Control colonies. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l8 level3 lfo10'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Started liquid cultures for O/N </p>
+<div align=center>
+<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+ style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
+ mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Piece</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Concentration (<span
+  class=SpellE>ng</span>/µl)</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Piece</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>Concentration (<span
+  class=SpellE>ng</span>/µl)</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE>TetR</span>
+  Backbone A</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>148.4</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>E</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>75.9</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>B</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>123.5</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>F</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>76.7</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>C</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>109.1</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>G</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>78.0</p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes'>
+  <td width=120 valign=top style='width:119.7pt;border:solid windowtext 1.0pt;
+  border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>D</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>29.3</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>H</p>
+  </td>
+  <td width=120 valign=top style='width:119.7pt;border-top:none;border-left:
+  none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+  <p class=MsoNormal style='line-height:normal'>74.3</p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'><span style="mso-spacerun:yes">&nbsp;</span></b></p>
+<a name="3"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 13. 6/16/2014 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level1 lfo11'><![if !supportLists]><span
+style='color:red'><span style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span style='color:red'>Plan to do:<o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level2 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Negative control: freeze and <span class=SpellE>miniprep</span>
+-&gt; Send to Medical School for sequencing</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level2 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Positive control: One-pot -&gt; Transform/plate</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level1 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>:</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level2 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Freeze and <span class=SpellE>miniprep</span> negative
+control</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level2 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><b style='mso-bidi-font-weight:normal'>Negative Control</b>:
+Ran Sequencing PCR for (-) <span class=SpellE>lacZ</span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level3 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Visualization gel (smear)</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level3 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Rerun sequencing PCR with new settings- failure</p>
+<p class=MsoNormal style='margin-left:1.0in;line-height:normal'><span
+class=SpellE>Redux</span> starting with DPN1 (-) <span class=SpellE>lacZ</span>
+#5 </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level3 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Purify</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level3 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=SpellE>Nanoquant</span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level3 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Blunt end ligation</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level3 lfo11'><![if !supportLists]><span
+style='color:#38761D'><span style='mso-list:Ignore'>&#9632;<span
+style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+style='color:#38761D'>Transformation/Electroporation and plating on <b
+style='mso-bidi-font-weight:normal'>cm</b> plates <o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level2 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><b style='mso-bidi-font-weight:normal'>Positive Control
+</b>(EYFP + <span class=SpellE>TetR</span> backbone) Run One Pot</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level3 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Transformed</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level3 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=GramE>plated</span> 100uL, rest at 8:05pm</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l13 level3 lfo11'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Incubate o/n</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 14. 6/17/2014 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l31 level1 lfo12'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Pick both
+Positive and Negative Control colonies</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l31 level2 lfo12'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Positive: <span class=SpellE><b style='mso-bidi-font-weight:
+normal'>kan</b></span><b style='mso-bidi-font-weight:normal'> antibiotic</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l31 level2 lfo12'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Negative: <b style='mso-bidi-font-weight:normal'>cm
+antibiotic</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l31 level3 lfo12'><![if !supportLists]><span
+style='color:red'><span style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span class=GramE><b style='mso-bidi-font-weight:
+normal'><span style='color:red'>no</span></b></span><b style='mso-bidi-font-weight:
+normal'><span style='color:red'> colonies seen as of 10 AM<o:p></o:p></span></b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l31 level3 lfo12'><![if !supportLists]><span
+style='color:red'><span style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span class=GramE><b style='mso-bidi-font-weight:
+normal'><span style='color:red'>colonies</span></b></span><b style='mso-bidi-font-weight:
+normal'><span style='color:red'> seen at 12 noon! <o:p></o:p></span></b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l31 level1 lfo12'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l31 level2 lfo12'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=GramE>redesigned</span> new PBJ003 plasmid
+map and designed/verified new sequencing primers. Sent them to Cheryl for
+verification. SAB primers have not yet arrived. Liquid cultures into the
+incubator by 9PM</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 15. 6/18/14 (Wednesday) -Worldly Wednesday!</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level1 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>:</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level2 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Get both <span class=SpellE>Pos</span> and <span
+class=SpellE>Neg</span> frozen and <span class=SpellE>miniprepped</span> -&gt;
+send off for sequencing</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level3 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=GramE>concurrently</span> with SAB?</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level2 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Go to <span class=SpellE>Worldy</span> Wednesday Lunch
+hosted by Andrew Ng </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level2 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Work on SAB?</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level1 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level2 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Froze both <span class=SpellE>Pos</span> and <span
+class=SpellE>Neg</span> and <span class=SpellE>miniprepped</span>. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level3 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=SpellE>Pos</span> process went to
+sequencing. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level3 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Negative control: start at <span class=SpellE>seq</span>
+PCR. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level2 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Ran SAB <span class=SpellE>SeqPCR</span>. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level3 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=GramE>negative</span> results</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l21 level2 lfo13'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Benjamin Colony PCR negative control (- <span
+class=SpellE>lacZ</span>) from old plates</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=402
+ style='width:402.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=65 style='width:65.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Pos</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #1<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>54.1<o:p></o:p></span></p>
+  </td>
+  <td width=65 style='width:65.0pt;border-top:none;border-left:none;border-bottom:
+  solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Neg</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #1<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>92.5<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Pos</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #2<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>163.4<o:p></o:p></span></p>
+  </td>
+  <td width=65 style='width:65.0pt;border-top:none;border-left:none;border-bottom:
+  solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Neg</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #2<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>73.1<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Pos</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #3<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>63.7<o:p></o:p></span></p>
+  </td>
+  <td width=65 style='width:65.0pt;border-top:none;border-left:none;border-bottom:
+  solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Neg</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #3<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>78.8<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Pos</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #4<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>109.5<o:p></o:p></span></p>
+  </td>
+  <td width=65 style='width:65.0pt;border-top:none;border-left:none;border-bottom:
+  solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Neg</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #4<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>126.5<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 16. 6/19/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level1 lfo14'><![if !supportLists]><span
+style='color:red'><span style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span style='color:red'>Plan to do: <o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level1 lfo14'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>:</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level2 lfo14'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Re-designed the SEQ primers for SAB. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level2 lfo14'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>CPCR for (-<span class=GramE>)<span class=SpellE>lacZ</span></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level3 lfo14'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=GramE>move</span> on to (-)cph8</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level2 lfo14'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=GramE>begin</span> work on PBJ003</p>
+<div align=center>
+<table class=MsoTableLightList border=1 cellspacing=0 cellpadding=0 width=527
+ style='width:527.0pt;border-collapse:collapse;border:none;mso-border-alt:solid black 1.0pt;
+ mso-border-themecolor:text1;mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:-1;mso-yfti-firstrow:yes;height:15.0pt'>
+  <td width=106 nowrap valign=top style='width:106.0pt;border-top:solid black 1.0pt;
+  mso-border-top-themecolor:text1;border-left:solid black 1.0pt;mso-border-left-themecolor:
+  text1;border-bottom:none;border-right:none;background:black;mso-background-themecolor:
+  text1;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-yfti-cnfc:5'><b><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman";color:white;mso-themecolor:background1'>Name<o:p></o:p></span></b></p>
+  </td>
+  <td width=421 nowrap valign=top style='width:421.0pt;border-top:solid black 1.0pt;
+  mso-border-top-themecolor:text1;border-left:none;border-bottom:none;
+  border-right:solid black 1.0pt;mso-border-right-themecolor:text1;background:
+  black;mso-background-themecolor:text1;padding:0in 5.4pt 0in 5.4pt;height:
+.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-yfti-cnfc:1'><b><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman";color:white;mso-themecolor:background1'>Primer<o:p></o:p></span></b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:0;height:15.0pt'>
+  <td width=106 nowrap valign=top style='width:106.0pt;border:solid black 1.0pt;
+  mso-border-themecolor:text1;border-right:none;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-yfti-cnfc:68'><span
+  class=SpellE><b><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>TetR_forward_new</span></b></span><b><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></b></p>
+  </td>
+  <td width=421 nowrap valign=top style='width:421.0pt;border:solid black 1.0pt;
+  mso-border-themecolor:text1;border-left:none;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-yfti-cnfc:64'><span
+  class=SpellE><span class=GramE><span style='font-size:12.0pt;font-family:
+  Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:"Times New Roman"'>tttttttgctcttcacacTCTCTACTAGAGTCACACTGGCTC</span></span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=106 nowrap valign=top style='width:106.0pt;border:none;border-left:
+  solid black 1.0pt;mso-border-left-themecolor:text1;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-yfti-cnfc:4'><span
+  class=SpellE><b><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>EYFP_forward_new</span></b></span><b><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></b></p>
+  </td>
+  <td width=421 nowrap valign=top style='width:421.0pt;border:none;border-right:
+  solid black 1.0pt;mso-border-right-themecolor:text1;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>tttttttgctcttca</span><span
+  style='font-size:12.0pt;font-family:"Lucida Grande";mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>attGGGAGCACTGTATTCACAGCGCTCCC</span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>tttaccggtctcaattcgcag</span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:15.0pt'>
+  <td width=106 nowrap valign=top style='width:106.0pt;border:solid black 1.0pt;
+  mso-border-themecolor:text1;border-right:none;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-yfti-cnfc:68'><b><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Spec2_Seq_For<o:p></o:p></span></b></p>
+  </td>
+  <td width=421 nowrap valign=top style='width:421.0pt;border:solid black 1.0pt;
+  mso-border-themecolor:text1;border-left:none;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-yfti-cnfc:64'><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>CGAGATAACCGTTGGCCTC<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:15.0pt'>
+  <td width=106 nowrap valign=top style='width:106.0pt;border:none;border-left:
+  solid black 1.0pt;mso-border-left-themecolor:text1;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-yfti-cnfc:4'><b><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>AraC2_Seq_For<o:p></o:p></span></b></p>
+  </td>
+  <td width=421 nowrap valign=top style='width:421.0pt;border:none;border-right:
+  solid black 1.0pt;mso-border-right-themecolor:text1;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>gttatccaataataactctgatagtgctag</span></span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes;height:15.0pt'>
+  <td width=106 nowrap valign=top style='width:106.0pt;border:solid black 1.0pt;
+  mso-border-themecolor:text1;border-right:none;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-yfti-cnfc:68'><b><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Bba2_Seq_Rev<o:p></o:p></span></b></p>
+  </td>
+  <td width=421 nowrap valign=top style='width:421.0pt;border:solid black 1.0pt;
+  mso-border-themecolor:text1;border-left:none;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal;mso-yfti-cnfc:64'><span
+  class=SpellE><span class=GramE><span style='font-size:12.0pt;font-family:
+  Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:"Times New Roman"'>cacttcactgacaccctcatc</span></span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 17. 6/20/14 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level1 lfo14'><![if !supportLists]><span
+style='color:red'><span style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span style='color:red'>Plan to do: <o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level1 lfo14'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>:</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level2 lfo14'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>PCR and gel extracted correct EYFP</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level2 lfo14'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=GramE>working</span> on getting piece <span
+class=SpellE>Ori</span> PCR -&gt; gel extract and setting up one-pot for PBJ003</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level2 lfo14'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>(-) <span class=GramE>cph8</span> took all the way
+through DNA clean &amp; concentrate and added wrong wash buffer</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l37 level3 lfo14'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9632;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>5µl purified DNA remaining in -20 (inside cph8 box)</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 18. 6/21/14 (Saturday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l41 level1 lfo15'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>:</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l41 level1 lfo15'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>:</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l41 level2 lfo15'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Negative Control<span class=GramE>:DPN1</span>, purify,
+<span class=SpellE>nanoquant</span> measure, blunt end ligation (BEL) and
+transform/plate on <b style='mso-bidi-font-weight:normal'>cm</b> </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l41 level2 lfo15'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>PBJ003: PCR -&gt; DNA Recover and <span class=SpellE>Nanoquant</span>
+measure. Digestion/Ligation (One-Pot) all pieces. Transformation/Plating on <b
+style='mso-bidi-font-weight:normal'>cm</b></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 19. 6/22/14 (Sunday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l7 level1 lfo16'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Pick
+colonies start liquid colonies for -cph8, pbj003</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l7 level1 lfo16'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Pick colonies
+start liquid colonies for -cph8, pbj003</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="4"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 20. 6/23/14 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l34 level1 lfo17'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Freeze, <span
+class=SpellE>miniprep</span>, sequencing <span class=SpellE>pcr</span>,
+visualization gel, send for sequencing</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l34 level1 lfo17'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: negative
+control, pbj003: Freeze, <span class=SpellE>miniprep</span>, sequencing <span
+class=SpellE>pcr</span>, visualization gel, send for sequencing</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l34 level2 lfo17'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>SAB: Sequencing PCR, visualization gel</p>
+<div align=center>
+<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0 width=402
+ style='width:402.0pt;border-collapse:collapse;border:none;mso-border-alt:solid windowtext 1.0pt;
+ mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt;mso-border-insideh:
+.0pt solid windowtext;mso-border-insidev:1.0pt solid windowtext'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border:solid windowtext 1.0pt;border-left:
+  none;mso-border-left-alt:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=65 style='width:65.0pt;border:solid windowtext 1.0pt;border-left:
+  none;mso-border-left-alt:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border:solid windowtext 1.0pt;border-left:
+  none;mso-border-left-alt:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Neg</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #1<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>128<o:p></o:p></span></p>
+  </td>
+  <td width=65 style='width:65.0pt;border-top:none;border-left:none;border-bottom:
+  solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;mso-border-top-alt:
+  solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ003 #1<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>146.3<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Neg</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #2<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>81.2<o:p></o:p></span></p>
+  </td>
+  <td width=65 style='width:65.0pt;border-top:none;border-left:none;border-bottom:
+  solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;mso-border-top-alt:
+  solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ003 #2<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>86.9<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Neg</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #3<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>47.8<o:p></o:p></span></p>
+  </td>
+  <td width=65 style='width:65.0pt;border-top:none;border-left:none;border-bottom:
+  solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;mso-border-top-alt:
+  solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ003 #3<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>69.4<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;
+  height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Neg</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #4<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>50.7<o:p></o:p></span></p>
+  </td>
+  <td width=65 style='width:65.0pt;border-top:none;border-left:none;border-bottom:
+  solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;mso-border-top-alt:
+  solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ003 #4<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext 1.0pt;mso-border-left-alt:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>95.2<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 21. 6/24/14 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l10 level1 lfo18'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>design
+experimental methods and procedures, design around hybrid promoter, find <span
+class=SpellE>lacI</span> resistant strain</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l10 level1 lfo18'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l10 level2 lfo18'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span class=GramE>did</span> modified SEQPCR on SAB and
+ran visualization gel- confirmed S and B present, A not present</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l10 level2 lfo18'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]>Ran PCR on part <span class=SpellE>AraC</span>/p15a <span
+class=GramE>( part</span> A), extracted and DNA recovered along with <span
+class=SpellE>nanoquant</span> measurements. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l10 level2 lfo18'><![if !supportLists]><span
+style='color:#38761D'><span style='mso-list:Ignore'>&#9675;<span
+style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+style='color:#38761D'>S<span class=GramE>,A,B</span> Digestion<o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l10 level2 lfo18'><![if !supportLists]><span
+style='color:#38761D'><span style='mso-list:Ignore'>&#9675;<span
+style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+style='color:#38761D'>S+A+B Ligation into <b style='mso-bidi-font-weight:normal'>SAB2</b><o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:1.0in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l10 level2 lfo18'><![if !supportLists]><span
+style='color:blue'><span style='mso-list:Ignore'>&#9675;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span></span><![endif]><span style='color:blue'>Transformation and
+Plating of <b style='mso-bidi-font-weight:normal'>SAB2 (failed)</b><o:p></o:p></span></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 22. 6/25/14 (Worldly Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l20 level1 lfo19'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Transform
+SAB2 C and D. Check sequencing results for the negative control and PBJ003 if
+they come in today. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l20 level1 lfo19'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: <span
+class=SpellE>Verfied</span> sequences for some positive control #4, some
+PBJ003. Sent T2 and E4 of PBJ003 in for sequencing confirmation, send in (-) <span
+class=SpellE>lacZ</span> sequencing on (-<span class=GramE>)cph8</span> plasmid,
+transformed and plated SAB2 C and D. Design primers for hybrid promoter</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 23. 6/26/14 (Thursday)<o:p></o:p></b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Pick and
+grow colonies for liquid culture</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done: </span>Grew
+liquid Colonies for SAB2. Work on website. Look up experimental procedures</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 24. 6/27/14 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Freeze and <span
+class=SpellE>miniprep</span> SAB2; Sequencing PCR, Send for sequencing</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done: </span>Freeze and
+<span class=SpellE>miniprep</span> SAB2; Sequencing PCR, <span
+style='color:red'>PCR fail, will re run with a gradient. </span>Verified
+completely PBJ003 #2, #4. (-) <span class=SpellE><span class=GramE>lacZ</span></span>
+funky. Received primers for hybrid promoter</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=437
+ style='width:437.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>SAB #1<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>86.6<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>SAB #2<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>97.1<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>SAB #3<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>79.1<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>SAB #4<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>65.8<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'><o:p>&nbsp;</o:p></b></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'><o:p>&nbsp;</o:p></b></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 25. 6/28/14 <span class=GramE>( Saturday</span>)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Run
+Sequencing PCR (gradient) for SAB2C #2. Send out purified PCR product for
+Sequencing. <span class=GramE>Set up experimental procedures</span>, <span
+class=GramE>write out experiment protocol</span>.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done: </span>Ran
+sequencing PCR with a gradient for SAB2 C#2. Verified negative control
+completely finished. </p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 26. 6/29 /14 <span class=GramE>( Sunday</span>)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Run SEQPCR
+(gradient) for SAB2C #2 and visualize. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done: </span>Ran SEQPCR
+for SAB2C #1. Same results on from 6/28/14. Redesigned primers for new plasmid
+(not using SAB anymore) called <span class=SpellE>pHOP</span></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="5"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 27. 6/30 /14 <span class=GramE>( Monday</span>)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: <o:p></o:p></span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done: </span>picked ice
+mix from Ray’s pSB1C3-BBa_M30109 frozen stock. Preparing for <span
+class=SpellE>miniprep</span> tomorrow morning (0800). </p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 28. 7/1 /14 <span class=GramE>( Tuesday</span>)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span><span
+class=SpellE>Miniprep</span> pSB1C3-BBa_M30109. Do a digestion and gel
+visualization step for pSB1C3-BBa_M30109 to see if <span class=GramE>its</span>
+legit. Start PCR and steps for <span class=SpellE>PBJ_Hybrid</span> (PBJ003 +
+hybrid promoter)</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l4 level1 lfo20'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done: </span>Saw
+nothing in the first digestion, could not re-run digestion due to no enzyme
+left. PCR, DPN1, Clean, BEL of PBJ_Hyb_001, transform, plate</p>
+<div align=center>
+<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0 width=527
+ style='width:527.0pt;margin-left:4.65pt;border-collapse:collapse;border:none;
+ mso-border-alt:solid black .5pt;mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt;
+ mso-border-insideh:.5pt solid black'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:15.0pt'>
+  <td width=106 nowrap valign=bottom style='width:106.0pt;border:solid black 1.0pt;
+  border-right:none;mso-border-top-alt:solid black .5pt;mso-border-left-alt:
+  solid black .5pt;mso-border-bottom-alt:solid black .5pt;background:black;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Name<o:p></o:p></span></b></p>
+  </td>
+  <td width=421 nowrap valign=bottom style='width:421.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;background:black;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Primer<o:p></o:p></span></b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:none;border-left:solid black 1.0pt;
+  border-bottom:solid black 1.0pt;border-right:none;mso-border-top-alt:solid black .5pt;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJ_hyb_for</span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=421 nowrap style='width:421.0pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black .5pt;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ <span class=SpellE><i>cattatacgagccggatgattaattgtcaa</i></span>
+  <span class=SpellE><b>ttt</b></span> CATTAAAAATGCGATCCTAAC 3’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:none;border-left:solid black 1.0pt;
+  border-bottom:solid black 1.0pt;border-right:none;mso-border-top-alt:solid black .5pt;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJ_hyb_rev</span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=421 nowrap style='width:421.0pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black .5pt;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ <span class=SpellE><i>tgtggaattgtgagcggataacaatttcacaca</i></span><i>
+  </i>ATGATAGATCCCGTCGTTTTAC 3’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:15.0pt'>
+  <td width=106 nowrap valign=bottom style='width:106.0pt;border-top:none;
+  border-left:solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:
+  none;mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-left-alt:solid black .5pt;mso-border-bottom-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'></td>
+  <td width=421 nowrap valign=bottom style='width:421.0pt;border-top:none;
+  border-left:none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-top-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'></td>
+ </tr>
+ <tr style='mso-yfti-irow:4;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:none;border-left:solid black 1.0pt;
+  border-bottom:solid black 1.0pt;border-right:none;mso-border-top-alt:solid black .5pt;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJ_hyb_seq_for</span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=421 nowrap style='width:421.0pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black .5pt;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ GCGACTTGATGCTCTTGATC 3’<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5;mso-yfti-lastrow:yes;height:15.0pt'>
+  <td width=106 nowrap style='width:106.0pt;border-top:none;border-left:solid black 1.0pt;
+  border-bottom:solid black 1.0pt;border-right:none;mso-border-top-alt:solid black .5pt;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  mso-border-bottom-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJ_hyb_seq_rev</span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=421 nowrap style='width:421.0pt;border-top:none;border-left:none;
+  border-bottom:solid black 1.0pt;border-right:solid black 1.0pt;mso-border-top-alt:
+  solid black .5pt;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>5’ CGAGGTCATATTCCAGTCG 3’<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='margin-left:.5in;line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 29. 7/2/14 (Worldly Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l15 level1 lfo21'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>make LB/cm
+plates (follow protocol), look at plated cultures and wrap when necessary. <span
+class=SpellE><span class=GramE>pHOP</span></span> primers should arrive today
+so we can get started working on that</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l15 level1 lfo21'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Pulled
+plates (2) from incubator, wrapped them and put into 4 <span class=GramE>fridge</span>.
+Picked colonies for PBJ_hyb_001. <span class=GramE>started</span> <span
+class=SpellE>pHOP</span> procedures. </p>
+<p class=MsoNormal style='margin-left:18.05pt;line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='margin-left:18.05pt;line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 30. 7/3/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l38 level1 lfo22'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span><span
+class=SpellE>Miniprep</span> -&gt; <span class=GramE>send</span> off for
+sequencing for PBJ_hyb_001. One pot then Transform and plate <span
+class=SpellE>pHOP</span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l38 level1 lfo22'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: <span
+class=SpellE>Miniprep</span>-&gt; visualize for PBJ_hyb_001. Plan on preparing
+for sequencing on Sunday. One <span class=GramE>pot,</span> transformed and
+plated <span class=SpellE>pHOP</span>. </p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=437
+ style='width:437.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ_hyb</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  E1<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>187.3<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ_hyb</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  H1<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>204<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ_hyb</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  E2<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>90<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ_hyb_H2<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>160.3<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 31. 7/4/14 (Friday. ‘<span class=SpellE>Murica</span>.)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l18 level1 lfo23'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: Pick <span
+class=SpellE>pHOP</span> colonies and start liquid cultures with <span
+class=SpellE><span class=GramE>kan</span></span> for O/N.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l18 level1 lfo23'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Start
+liquid cultures</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 32. 7/5/14 (Saturday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l40 level1 lfo24'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: Freeze -&gt;
+Run SEQPCR Gel on <span class=SpellE>pHOP</span>.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l40 level1 lfo24'><![if !supportLists]><span
+style='color:#38761D'><span style='mso-list:Ignore'>&#9679;<span
+style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+style='color:#38761D'>Got done</span>: <span class=SpellE>pHOP</span> gel bands
+are of the wrong length. <span style='color:#38761D'><o:p></o:p></span></p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=437
+ style='width:437.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>pHOP</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> 1<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>76.4<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>pHOP</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> 3<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>157.9<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>pHOP</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> 2<o:p></o:p></span></p>
+  </td>
+  <td width=120 style='width:120.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>128<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>pHOP</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> 4<o:p></o:p></span></p>
+  </td>
+  <td width=114 style='width:114.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>164.8<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 33. 7/6/14 (Sunday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l2 level1 lfo25'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: Get sequencing
+ready for PBJ_hyb_001, re run <span class=SpellE>pHOP</span> SEQPCR</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l2 level1 lfo25'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Go
+sequencing ready for hybrid promoter. <span class=SpellE><span class=GramE>pHOP</span></span>
+SEQPCR gel visualization not of correct length. </p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="6"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 34. 7/7/14 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l2 level1 lfo25'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: One Pot
+enhanced pHOP2/ Colony PCR pHOP3</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l2 level1 lfo25'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Colony PCR
+for pHOP3 showed wrong bands. Enhanced One Pot for pHOP2, transformed, plated
+on LB/<span class=SpellE>Kan</span></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 35. 7/8/14 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l2 level1 lfo25'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: Start Liquid
+culture for pHOP2</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l2 level1 lfo25'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Started
+Liquid culture for pHOP2, Colony PCR pHOP2, Verified Sequencing for PBJ_hyb_001
+F, ready to go.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 36. 7/9/14 (Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l2 level1 lfo25'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: Sequencing <span
+class=SpellE>pcr</span> for pHOP2, send for sequencing</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l2 level1 lfo25'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Sequencing
+PCR failed, do a modified digest of BBa_M30109 using BSG1, failed. Work on
+degradation tag design (finished)</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 37. 7/10/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l32 level1 lfo26'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: Get a
+protocol written for testing of PBJ_hyb_001 (light and dark)</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l32 level1 lfo26'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Protocol
+for testing written out. Started liquid cultures in light/dark</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 38. 7/11/14 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l14 level1 lfo27'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>:</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l14 level1 lfo27'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Tested hybrid
+and PBJ003 without chromophore. <span class=GramE>designed</span> primers for
+PBJ004 (with <span class=SpellE>Ptet</span> promoter <span class=SpellE>Ptet-pp</span>+
+plasmid from Dr. Moon to control EYFP). </p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=428
+ style='width:428.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:15.0pt'>
+  <td width=65 nowrap valign=bottom style='width:65.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Name<o:p></o:p></span></b></p>
+  </td>
+  <td width=363 nowrap style='width:363.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Primer<o:p></o:p></span></b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:15.0pt'>
+  <td width=65 nowrap valign=bottom style='width:65.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>PBJ003_F<o:p></o:p></span></p>
+  </td>
+  <td width=363 nowrap style='width:363.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'><span style="mso-spacerun:yes">&nbsp;</span><span
+  class=SpellE><span class=GramE>atcagtgatagagatactgagcacatctcCGGATAACAATTTCACACATACTAGAG</span></span><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:15.0pt'>
+  <td width=65 nowrap valign=bottom style='width:65.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>PBJ003_R<o:p></o:p></span></p>
+  </td>
+  <td width=363 nowrap valign=bottom style='width:363.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>agggatgtcaatctctatcactgatagggaGACGAAAGGTGATACCGCTC</span></span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:15.0pt'>
+  <td width=65 nowrap valign=bottom style='width:65.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>SEQ004_F<o:p></o:p></span></p>
+  </td>
+  <td width=363 nowrap valign=bottom style='width:363.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GTGCCAAGGGAAAAACTAGTC<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes;height:15.0pt'>
+  <td width=65 nowrap valign=bottom style='width:65.0pt;border:solid black 1.0pt;
+  border-right:none;mso-border-top-alt:solid black .5pt;mso-border-left-alt:
+  solid black .5pt;mso-border-bottom-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>SEQ004_R<o:p></o:p></span></p>
+  </td>
+  <td width=363 nowrap valign=bottom style='width:363.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>gaacttcagggtcagcttg</span></span></span><span
+  style='font-size:12.0pt;font-family:Cambria;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'>&lt;<span class=GramE>insert</span>
+table 1&gt;</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="7"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 39. 7/14/14 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l16 level1 lfo28'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: Begin
+cloning for degradation tags on PBJ003 AND PBJ_hyb_001</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l16 level1 lfo28'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Started
+PCR for Degradation Tags as well as <span class=SpellE>Ptet-pp</span> (same as:
+<span class=SpellE>Ptet</span>-T*2) minus the <span class=SpellE>TetR</span>
+(as a control). Designed light-box. </p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 40. 7/15/14 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l28 level1 lfo29'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: meet with
+Andrew to go over light-box ordering. Continue cloning projects. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l28 level1 lfo29'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: cloning <span
+class=GramE>project</span> up to Purification after Dpn1- no DNA present so
+re-did from PCR. Design primers for <span class=SpellE>pChrom</span>.</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=306
+ style='width:4.25in;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:15.0pt'>
+  <td width=151 nowrap valign=bottom style='width:151.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Name<o:p></o:p></span></b></p>
+  </td>
+  <td width=155 nowrap valign=bottom style='width:155.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Primers<o:p></o:p></span></b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:15.0pt'>
+  <td width=151 nowrap valign=bottom style='width:151.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>pChrom</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>_seq_for</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=155 nowrap valign=bottom style='width:155.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GTGCCACCTGACGTCTAAG<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;mso-yfti-lastrow:yes;height:15.0pt'>
+  <td width=151 nowrap valign=bottom style='width:151.0pt;border:solid black 1.0pt;
+  border-right:none;mso-border-top-alt:solid black .5pt;mso-border-left-alt:
+  solid black .5pt;mso-border-bottom-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>pChrom</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>_seq_rev</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=155 nowrap valign=bottom style='width:155.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GCTGACTTGACGGGACTC<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 41. 7/16/14 (Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l6 level1 lfo30'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Keep cloning</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l6 level1 lfo30'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>kept working
+on <span class=GramE>pet(</span>-)<span class=SpellE>tetR</span>,
+PBJ003_DegTetR, and PB_hyb_001 <span class=SpellE>DegTetR</span> up to
+transformation and plating</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 42. 7/17/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l24 level1 lfo31'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: Check on and
+wrap plates. Start liquid cultures for both degradation tags, <span
+class=SpellE><span class=GramE>ptet</span></span><span class=GramE>(</span>-)<span
+class=SpellE>tetR</span> and <span class=SpellE>biobrick</span> part from
+shipped frozen stock.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l24 level1 lfo31'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Checked
+plates, wrapped them around 10:20 AM. Started Liquid culture at 6pm.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 43. 7/18/14 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l24 level1 lfo31'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: <span
+class=SpellE>Miniprep</span>, sequencing PCR degradation tags, <span
+class=SpellE><span class=GramE>ptet</span></span><span class=GramE>(</span>-).
+Start cloning for <span class=SpellE>pChrom</span>: <span class=SpellE>Miniprep</span>
+<span class=SpellE>Biobrick</span> part, digestion &amp; ligation alongside
+PFM109 &amp; transform</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l24 level1 lfo31'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Sequencing
+PCR for degradation tags. <span class=SpellE><span class=GramE>ptet</span></span>(-)
+<span class=SpellE>doesnt</span> look right. Only <span class=SpellE>miniprepped</span>
+<span class=SpellE>biobrick</span> part.</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=461
+ style='width:461.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:15.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;mso-border-alt:
+  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border:solid windowtext 1.0pt;border-left:
+  none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border:solid windowtext 1.0pt;border-left:
+  none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µ<span class=GramE>l)3</span><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:15.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Hyb</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  degradation 1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>82.3<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>hyb</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> <span class=SpellE>deg</span>
+<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>67.1<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:15.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>hyb</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> <span class=SpellE>deg</span>
+<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>143.4<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>hyb</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> <span class=SpellE>deg</span>
+<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>48<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:15.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ 003 1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>153<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ 003 2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>61.1<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;height:15.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ 003 3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>612.5<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ 003 4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>75.1<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5;height:15.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>ptet</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> (-) <span class=SpellE>tetR</span>
+<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>248.8<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>ptet</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> (-) <span class=SpellE>tetR</span>
+<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>386.4<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6;height:15.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>ptet</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> (-) <span class=SpellE>tetR</span>
+<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>67.6<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-bottom-alt:solid windowtext .5pt;mso-border-right-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>ptet</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> (-) <span class=SpellE>tetR</span>
+<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>37.7<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7;mso-yfti-lastrow:yes;height:15.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Bba_k1017726<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>86.2<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>pFM109</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>8.9<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 44. 7/19/14 (Saturday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l24 level1 lfo31'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do</span>: Sequencing
+grad PCR for <span class=SpellE><span class=GramE>ptet</span></span><span
+class=GramE>(</span>-), Sequential Digestion for pFM109, Bba_K1017726 -&gt; Gel
+Extract -&gt; Ligation -&gt; Transform &amp; plate</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l24 level1 lfo31'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:#38761D'>Got done</span>: Digested with
+EcoR1, Digested with <span class=SpellE>SpeI</span>, Ligation; Sequencing PCR
+for <span class=SpellE><span class=GramE>ptet</span></span><span class=GramE>(</span>-):
+piece size not as expected ran colony <span class=SpellE>pcr</span>- band
+length still indicates <span class=SpellE>tetR</span> present</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 45. 7/20/14 (Sunday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l9 level1 lfo32'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Prep <span
+class=SpellE>DegTag</span> products PBJ003 and hybrid for Sequencing. Transform
+and Plate <span class=SpellE>pChrom</span>.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l9 level1 lfo32'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Degradation
+Tag on <span class=SpellE>TetR</span> for both PBJ003 and hybrid are ready for
+sequencing. Transform and plate <span class=SpellE>pChrom</span> on <span
+class=SpellE>lb</span>/<span class=SpellE>kanR</span> plates. Put in 37 at
+:50pm</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="8"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 46. 7/21/14 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l42 level1 lfo33'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>send tags
+off for sequencing. Start <span class=SpellE><span class=GramE>ptet</span></span><span
+class=GramE>(</span>-) <span class=SpellE>tetR</span> cloning steps. Start
+liquid cultures for Chromophore project. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l42 level1 lfo33'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>tags sent
+for sequencing. <span class=GramE>finished</span> from PCR to dpn1 on <span
+class=SpellE>ptet</span> (-) <span class=SpellE>tetR</span>. <span class=GramE>colony</span>
+<span class=SpellE>pcr</span> for chromophore failed. Redo ligation and
+transformation</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 47. 7/22/14 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l30 level1 lfo34'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>redo colony <span
+class=SpellE>pcr</span> for chromophore. Work on <span class=SpellE><span
+class=GramE>ptet</span></span><span class=GramE>(</span>-)<span class=SpellE>tetR</span>
+positive control.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l30 level1 lfo34'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>kept working
+on <span class=SpellE><span class=GramE>ptet</span></span><span class=GramE>(</span>-)<span
+class=SpellE>tetR</span> positive control- dpn1 and <span class=SpellE>purify,transformed</span>
+and plated. Not enough colonies for chromophore to colony <span class=SpellE>pcr</span>.
+Started liquid culture at night.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 48. 7/23/14 (Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l43 level1 lfo35'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span><span
+class=SpellE>Miniprep</span> and sequencing <span class=SpellE>pcr</span> for
+chromophore. Check and wrap plates, start liquid cultures around 8 or 9. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l43 level1 lfo35'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span><span
+class=SpellE>Miniprep</span> and Sequencing PCR for chromophore. Prepped for <span
+class=SpellE>sequencingstarted</span> (-<span class=GramE>)<span class=SpellE>tetR</span></span>
+liquid cultures. <span class=GramE>worked</span> on PCR <span class=SpellE>seq</span>
+for Degradation tags on PBJ003 and <span class=SpellE>Phybrid</span>.<span
+style="mso-spacerun:yes">&nbsp; </span>“<span class=GramE>need</span>” to
+purify before sending <span class=SpellE>pcr</span> product. </p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=461
+ style='width:461.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>pChrom</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> #1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>142.2<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>pChrom</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> #2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>122.8<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>pChrom</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> #3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>112.8<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>pChrom</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> #4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>78.7<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 49. 7/24/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l43 level1 lfo35'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Purify PCR
+products. <span class=SpellE>Miniprep</span> and check sequencing <span
+class=SpellE>pcr</span> for (-<span class=GramE>)<span class=SpellE>tetR</span></span>.
+Send all for sequencing</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l43 level1 lfo35'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Purified PCR
+product for <span class=SpellE>chrom</span> and degradation tags. Prepped for
+sequencing. <span class=SpellE>Miniprep</span> and run sequencing PCR for (-<span
+class=GramE>)<span class=SpellE>tetR</span></span> (no bands)</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=461
+ style='width:461.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>(-) <span class=SpellE><span
+  class=GramE>tetR</span></span> 1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>102<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>(-) <span class=SpellE><span
+  class=GramE>tetR</span></span> 2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>129.6<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>(-) <span class=SpellE><span
+  class=GramE>tetR</span></span> 3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>455.3<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>(-) <span class=SpellE><span
+  class=GramE>tetR</span></span> 4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>906.4<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 50. 7/25/14 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l19 level1 lfo36'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Gradient
+SEQPCR on (-<span class=GramE>)<span class=SpellE>tetR</span></span>, run a gel
+<span class=SpellE>visualizatoin</span>, prep for sequencing.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l19 level1 lfo36'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Ran gradient
+on (-<span class=GramE>)<span class=SpellE>tetR</span></span> #3 and 4 with gel
+visualization, saw bands at wrong length. <span class=GramE>doing</span> colony
+PCR tomorrow. </p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 51. 7/26/14 (Saturday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l17 level1 lfo37'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Brute Force
+Colony PCR (basically picking 32 colonies!). Freeze and <span class=SpellE>Miniprep</span>
+correct ones. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l17 level1 lfo37'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Brute Force
+Colony PCR and gel visualization finished.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 52. 7/27/14 (Sunday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l26 level1 lfo38'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Start all
+cloning procedures by taking #4 of the degradation tags from hybrid and
+pBJ003.<span style="mso-spacerun:yes">&nbsp; </span>Prep for sequencing the <span
+class=SpellE>miniprepped</span><span style="mso-spacerun:yes">&nbsp; </span>(-<span
+class=GramE>)<span class=SpellE>tetR</span></span> DNA samples (#9,12,15,16). </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l26 level1 lfo38'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Prep (-<span
+class=GramE>)<span class=SpellE>tetR</span></span> for sequencing. Start
+cloning PBJ004 and PBJ_hyb_002</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="9"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 53. 7/28/14 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l23 level1 lfo39'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Digest <span
+class=SpellE>Biobrick</span> part to check for correct lengths… continue
+cloning of PBJ004 and PBJ_hyb_002 with the new <span class=SpellE>ptet</span>
+promoter insertion. Look more at LED grid coding. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l23 level1 lfo39'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Digestion of
+<span class=SpellE>biobrick</span> part only showed a smear. Still send both <span
+class=SpellE>miniprep</span> and <span class=SpellE>pcr</span> product for
+sequencing </p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 54. 7/29/14 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l29 level1 lfo40'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span><span
+style='mso-bidi-font-size:11.0pt;color:windowtext'>Check plates in the morning,
+wrap. Start cloning for PBJ005 and Hybrid <span class=GramE>003 which are
+PBJ003</span> and Hybrid001 </span><span style='mso-bidi-font-size:11.0pt;
+mso-fareast-font-family:"Times New Roman";color:windowtext;mso-bidi-font-weight:
+bold'>plasmids that just have the new <span class=SpellE>ptet</span> promoter
+inserted. </span>Start liquid cultures at night.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l29 level1 lfo40'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>plates
+checked and wrapped. Sanity check #2 for <span class=SpellE>biobrick</span>
+part, doing a double digestion. Started cloning of PBJ005 and Hybrid003. Liquid
+cultures started. Retransform003, <span class=GramE>order new sequencing
+primer, redo</span> double digestion.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 55. 7/30/14 (Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Freeze, <span
+class=SpellE>miniprep</span>, <span class=GramE>send</span> off for sequencing
+of PBJ004 and Hybrid002. Delete the CAC in the maps for PBJ003 with <span
+class=SpellE>DegTagTetR</span> and <span class=SpellE>PBJ_hyb</span> with <span
+class=SpellE>DegTagTetR</span>. Get iGEM calendar set up. Email <span
+class=SpellE>Pakrasi</span> and Moon about setting up final iGEM meeting</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Start
+Modeling of light promoter system. (-) <span class=SpellE><span class=GramE>tetR</span></span>
+still no luck.<span style="mso-spacerun:yes">&nbsp; </span>Sequence verified <span
+class=SpellE>pCHROM</span> #1. Freeze <span class=SpellE>miniprep</span>,
+PBJ004, Hybrid 002. Start liquid cultures for PBJ005, Hybrid 003.</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=461
+ style='width:461.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ004 #1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>145.6<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ004 #2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>165.4<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ004 #3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>97.7<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ004 #4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>122.3<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Hybrid002 #1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>80<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Hybrid002 #2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>436.8<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Hybrid002 #3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>102.4<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Hybrid002 #4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>117.5<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 56. 7/31/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Make LB/
+(spec + cm) &amp; (spec + <span class=SpellE>kan</span>) plates. <span
+class=SpellE>Seq</span> PCR for (-) <span class=SpellE>tetR</span>. Freeze <span
+class=SpellE>miniprep</span> PBJ005, Hybrid003. Send PBJ004, PBJ005, Hybrid
+, Hybrid 002, (-) <span class=SpellE>tetR</span> for sequencing. Design
+experimental protocol. Transform.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Made plates.
+<span class=SpellE>Miniprep</span> PBJ005, hybrid 003. Sequencing PCR, gel
+visualization for PBJ005, hybrid003, (-) <span class=SpellE><span class=GramE>tetR</span></span>.
+<span class=GramE>Gel visualize</span>. Gradient PCR hybrid003, (-<span
+class=GramE>)<span class=SpellE>tetR</span></span>, visualize. Found Hybrid003
+E, 1F, (-<span class=GramE>)<span class=SpellE>tetR</span></span> wrong bands.
+Prep PBJ004, PBJ005, <span class=GramE>hybrid002</span> for sequencing.
+Transform/ plate <span class=SpellE>pchrom</span> + (<span class=SpellE>pos</span>/<span
+class=SpellE>neg</span>/pbj003<span class=GramE>,phybrid</span>).</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=461
+ style='width:461.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ005 #1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>145.6<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ005 #2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>165.4<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ005 #3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>97.7<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>PBJ005 #4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>122.3<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Hybrid003 #1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>80<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Hybrid003 #2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>436.8<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Hybrid003 #3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>102.4<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Hybrid003 #4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>117.5<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 57. 8/1/14 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Prep
+Hybrid003 for sequencing. Send all in before 9. Pick <span class=SpellE>cotransformed</span>
+colonies. Start liquid culture.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Sent for
+sequencing. Colonies not growing well. Colony PCR pchrom3 (from 7/29/14). Work
+on modeling.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 58. 8/2/14 (Saturday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Check
+plates. Wrap. Start liquid cultures</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Wrap colony
+PCR plate. <span class=SpellE>Transformants</span> not growing. ***
+Realization, <span class=SpellE>pCHROM</span> has <span class=SpellE><span
+class=GramE>kan</span></span> resistance, so using the wrong plates the whole
+time. Started liquid cultures for pCHROM3, pCHROM1.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 59. 8/3/14 (Sunday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span><span
+class=SpellE>Miniprep</span> pCHROM3 and 1</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Freeze
+pCHROM3, <span class=SpellE>Miniprep</span> pCHROM3 and 1. Sequencing results
+for new <span class=SpellE>Ptet</span> bad</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=461
+ style='width:461.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:
+  "Times New Roman"'>pChrom</span></span></span><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'> 1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>157.5<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>pChrom3</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>133.6<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>pChrom3</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #5<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>102.6<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>pChrom3</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #12<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>79.4<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=GramE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>pChrom3</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #13<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>91.5<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>&nbsp;<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>&nbsp;<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="10"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 60. 8/4/14 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Make <span
+class=SpellE>Kan</span>/CM plates. Check Sequence for pCHROM3. Transform and
+plate for experiment</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Made plates.
+Sequence PCR #1 showed multiple bands. Sequence PCR #2 showed wrong bands.
+Started new <span class=SpellE>Ptet</span> stuff over again from PCR.
+Transformed and plate experimental stuff</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 61. 8/5/14 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Wrap Plates.
+<span class=SpellE>Miniprep</span> PBJ003, PBJ_Hyb_001. Redo cloning for new <span
+class=SpellE>Ptet</span> plasmids. Start liquid cultures for experimentation.
+Work on presentation</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Wrap plates.
+<span class=SpellE>Miniprep</span>. Redo PBJ005</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 62. 8/6/14 (Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Run
+experiment. Induce. Check data</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Lighted
+growth chamber in use at 30 degrees. Cheryl advise to abort <span class=SpellE>util</span>
+tomorrow… Work on presentation. Purify, Transform PBJ005 redo. Design <span
+class=SpellE>PBJpos</span></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 63. 8/7/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Run
+experiment. Induce. Check data</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Ran
+experiment. Dilute to OD .1, grow for 2 hours, induction. Start PBJ005 liquid
+cultures, measure fluorescence. NO FLUORESCENCE. Plan out new positive control,
+knocking out <span class=SpellE>CcaR</span>/<span class=SpellE>CcaS</span>/Pcpcg2/<span
+class=SpellE>tetR</span> from PBJ003.</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=261
+ style='width:261.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:15.0pt'>
+  <td width=66 nowrap valign=bottom style='width:66.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Name<o:p></o:p></span></b></p>
+  </td>
+  <td width=195 nowrap valign=bottom style='width:195.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Primer<o:p></o:p></span></b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:15.0pt'>
+  <td width=66 nowrap valign=bottom style='width:66.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJpos_F</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=195 nowrap valign=bottom style='width:195.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GAGCGGTATCACCTTTCGTC<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:15.0pt'>
+  <td width=66 nowrap valign=bottom style='width:66.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJpos_R</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=195 nowrap valign=bottom style='width:195.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>CCTCGAGCTTAGGACGTCG<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:15.0pt'>
+  <td width=66 nowrap valign=bottom style='width:66.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>SEQpos_F</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=195 nowrap valign=bottom style='width:195.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GTTCTTTACGATGCCATTGG<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes;height:15.0pt'>
+  <td width=66 nowrap valign=bottom style='width:66.0pt;border:solid black 1.0pt;
+  border-right:none;mso-border-top-alt:solid black .5pt;mso-border-left-alt:
+  solid black .5pt;mso-border-bottom-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>SEQpos_R</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=195 nowrap valign=bottom style='width:195.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Cambria;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>CACCATCTAGTATTTCTCCTCTTTC<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'>&lt;<span class=GramE>data</span>
+table 2&gt;</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 64. 8/8/14 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do: </span>Send PBJ005
+for sequencing</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Froze, <span
+class=SpellE>miniprep</span>, Sequencing PCR. Send Purified <span class=SpellE>Seq</span>
+PCR product for sequencing. Jeffrey’s last day</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="11"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 65. 8/11/14 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Start
+cloning for <span class=SpellE>PBJpos</span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>PCR,
+visualize, DPN1, purify, Blunt end ligate. Transform <span class=SpellE>PBJpos</span>.
+Also Transform PSL2264</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 66. 8/12/14 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Start liquid
+culture for <span class=SpellE>PBJpos</span>, PSL2264</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Wrapped
+plates. Checked sequence results for PBJ005=bad. Started liquid cultures for <span
+class=SpellE>PBJpos</span></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 67. 8/13/14 (Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> <span
+class=SpellE>Miniprep</span>, send for sequencing, start liquid cultures for
+experimentation</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span><span
+class=SpellE>Miniprepped</span> and sent <span class=SpellE>PBJpos</span> #1-#4
+for sequencing. Started liquid cultures for pSL2264, <span class=SpellE>PBJpos</span>
+-4, current positive control and negative control for experimentation
+tomorrow.</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=461
+ style='width:461.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJpos</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>360.5<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJpos</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>349.9<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJpos</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>443.9<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJpos</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>270.8<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 68. 8/14/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Run
+fluorescence tests for old and new positive controls. Sanity check for pSL2264</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>pSL2264 not
+fluorescing as expected. <span class=SpellE>PBJpos</span> slightly better, <span
+class=GramE>Old</span> positive control and negative control as expected.
+Induce pSL2264 with IPTG. Design new PBJ003 with deletion of competing TSS</p>
+<p class=MsoNormal style='line-height:normal'>&lt;<span class=GramE>data</span>
+table 3&gt;</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 69. 8/15/14 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Design
+primers for new PBJ plasmids with modified RBS. Plan out fall semester work
+schedule.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l22 level1 lfo41'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Design
+primers for new PBJ plasmids with modified RBS.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="12"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 70. 8/25/14 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Start
+cloning for <span class=SpellE>PBJposrbs</span> (new positive control with
+enhanced RBS) until transformation</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>PCR, <span
+class=GramE>gel visualize</span>, DPN1, Ligation, Transformation, Plating. Work
+on Protocol section for website.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 71. 8/26/14 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Start Liquid
+Culture</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Start liquid
+culture. Work on protocols.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 72. 8/27/14 (Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> <span
+class=SpellE>Miniprep</span>. Run sequencing PCR, send for sequencing</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span><span
+class=SpellE>Miniprep</span>. Run sequencing PCR, results bad. Rerun sequencing
+PCR with correct primers.</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=461
+ style='width:461.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJpos_rbs</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>166.3<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJpos_rbs</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>165.2<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJpos_rbs</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>107.9<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJpos_rbs</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>271.5<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 73. 8/28/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Visualize
+sequencing PCR. PCR purify, prep for sequencing. Send</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>visualize
+sequencing PCR. PCR purify, prep for sequencing. Work on title and abstract</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 74. 8/29/14 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Send in for
+sequencing (morning). Make <span class=SpellE>electrocompetent</span> cells</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Send in for
+sequencing. Make <span class=SpellE>electrocompetent</span> cells. Work on
+title and abstract.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="13"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 75. 9/1/14 (Monday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Sequence <span
+class=GramE>verify</span> <span class=SpellE>PBJpos_RBS</span>. Start liquid
+cultures for experimentation.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Sequence <span
+class=GramE>verify</span> <span class=SpellE>PBJpos_RBS</span>. Start liquid
+cultures for experimentation. Final safety forms due<span class=GramE>.!</span></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 76. 9/2/14 (Tuesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Test for
+Fluorescence of newest positive control. </p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l1 level1 lfo42'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>tested for
+fluorescence. Dilute to 0.1 grew an extra 6 hours, tested again. EYFP did not
+fluoresce as expected. <span class=SpellE>Compard</span> 5’ UTRs between
+different positive controls. <span class=GramE>redesign</span> primers in order
+to change <span class=SpellE>pTet</span> from tabor to moon’s.</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=488
+ style='width:488.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:15.0pt'>
+  <td width=135 nowrap valign=bottom style='width:135.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Name<o:p></o:p></span></b></p>
+  </td>
+  <td width=353 nowrap valign=bottom style='width:353.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Primer<o:p></o:p></span></b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:15.0pt'>
+  <td width=135 nowrap valign=bottom style='width:135.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJptet_F</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=353 nowrap valign=bottom style='width:353.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>tagagatactgagcacatctcCGGATAACAATTTCACACATACTAGAG</span></span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:15.0pt'>
+  <td width=135 nowrap valign=bottom style='width:135.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Pbjptet_R</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=353 nowrap valign=bottom style='width:353.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>tcactgatagggatgtcaatctctatcactgatagggaACGAAAGGTGATACCGCTCG</span></span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:15.0pt'>
+  <td width=135 nowrap valign=bottom style='width:135.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJpos_seq_F</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=353 nowrap valign=bottom style='width:353.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Use PBJRBS <span class=SpellE>seq_F</span><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;height:15.0pt'>
+  <td width=135 nowrap valign=bottom style='width:135.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJpos_seq_R</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=353 nowrap valign=bottom style='width:353.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Use PBJRBS <span class=SpellE>seq_R</span><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5;height:15.0pt'>
+  <td width=135 nowrap valign=bottom style='width:135.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>BbaMoonptet_F</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=353 nowrap valign=bottom style='width:353.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Use <span class=SpellE>PBJptet_F</span><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6;height:15.0pt'>
+  <td width=135 nowrap valign=bottom style='width:135.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>BBaMoonptet_R</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=353 nowrap valign=bottom style='width:353.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>tcactgatagggatgtcaatctctatcactgatagggaAACTGCTTTCGGTTAGACTTC</span></span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7;height:15.0pt'>
+  <td width=135 nowrap valign=bottom style='width:135.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>BbaMoon_seq_f</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=353 nowrap valign=bottom style='width:353.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>AACCGGTAACAGTACAAATCC<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:8;mso-yfti-lastrow:yes;height:15.0pt'>
+  <td width=135 nowrap valign=bottom style='width:135.0pt;border:solid black 1.0pt;
+  border-right:none;mso-border-top-alt:solid black .5pt;mso-border-left-alt:
+  solid black .5pt;mso-border-bottom-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>BbaMoon_seq_r</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=353 nowrap valign=bottom style='width:353.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Use PBJRBS <span class=SpellE>seq_R</span><o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'>&lt;Data table 4&gt;</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 77. 9/4/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Start
+cloning once primers <span class=GramE>are</span> received.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Organize
+primers for assembly. Start cloning for swapping <span class=SpellE>Ptet</span>
+on positive controls.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 78. 9/5/14 (Friday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Take cloning
+through transformation</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Gel,
+Digestion, Purification, Ligation, Transformation</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 79. 9/6/14 (Saturday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Start liquid
+<span class=SpellE>cultues</span></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Wrapped
+plates in the morning. Start liquid cultures at night</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 80. 9/7/14 (Sunday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> <span
+class=SpellE>Miniprep</span>, Sequencing PCR, Prep for sequencing.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span><span
+class=SpellE>Miniprep</span>, Sequencing PCR, Prepare for sequencing and send
+off tomorrow morning.</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=461
+ style='width:461.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;padding:
+in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Piece<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border:solid windowtext 1.0pt;border-left:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='mso-bidi-font-size:
+.0pt;mso-fareast-font-family:"Times New Roman"'>Concentration (<span
+  class=SpellE>ng</span>/µl)<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJposmoon</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>82.9<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJposmoon</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>179.4<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJposmoon</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>91.1<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>PBJposmoon</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>50.5<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Bbaposmoon</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #1<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>84.9<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Bbaposmoon</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #2<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>180.3<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes;height:16.0pt'>
+  <td width=103 style='width:103.0pt;border:solid windowtext 1.0pt;border-top:
+  none;padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Bbaposmoon</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #3<o:p></o:p></span></p>
+  </td>
+  <td width=126 style='width:1.75in;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>72.7<o:p></o:p></span></p>
+  </td>
+  <td width=100 style='width:100.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>Bbaposmoon</span></span><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>
+  #4<o:p></o:p></span></p>
+  </td>
+  <td width=132 style='width:132.0pt;border-top:none;border-left:none;
+  border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
+  padding:0in 5.4pt 0in 5.4pt;height:16.0pt'>
+  <p class=MsoNormal align=right style='text-align:right;line-height:normal'><span
+  style='mso-bidi-font-size:11.0pt;mso-fareast-font-family:"Times New Roman"'>28.5<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<a name="14"></a>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 81. 9/10/14 (Wednesday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Check on
+sequences</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span>Verified
+sequence for <span class=SpellE>Bba_pos_moon</span>. Started liquid cultures
+for experimentation tomorrow. Start designing primers for <span class=SpellE>biobricking</span>.</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><b style='mso-bidi-font-weight:
+normal'>Day 82. 9/11/14 (Thursday)</b></p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:red'>Plan to do:</span> Check for
+fluorescence of new <span class=SpellE>pTet</span>. Finish and clarify design
+of primers for <span class=SpellE>biobricking</span>.</p>
+<p class=MsoNormalCxSpMiddle style='margin-left:.5in;mso-add-space:auto;
+text-indent:-17.95pt;line-height:normal;mso-list:l36 level1 lfo43'><![if !supportLists]><span
+style='mso-list:Ignore'>&#9679;<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></span><![endif]><span style='color:green'>Got done: </span><span
+class=SpellE>Bba_pos_moon</span> fluoresces as expected!!!!! Designed primers
+to start <span class=SpellE>biobricking</span>. Sent for ordering.</p>
+<div align=center>
+<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width=487
+ style='width:487.0pt;margin-left:4.65pt;border-collapse:collapse;mso-yfti-tbllook:
+;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+ <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Name<o:p></o:p></span></b></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  background:black;padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><b><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman";color:white'>Primers<o:p></o:p></span></b></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:1;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Bba_PBJ_f</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>gaattcgcggccgcttctagagAGCCCATTGTGCTTTTC</span></span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:2;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Bba_PBJ_r</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>ctgcagcggccgctactagtaGATTATTAGGACCCACTTTCAC</span></span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:3;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJ_seq_prefix_f</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>CCTATAAAAATAGGCGTATCAC<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:4;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJ_seq_prefix_r</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GCAAAAGTGAGTATGGTG<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:5;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJ_seq_Suffix_f</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>CTTTGAGTGAGCTGATACC<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:6;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>PBJ_seq_Suffix_r</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>GATTAACAGCGCATTAGAG<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:7;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Bba_pos_f</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>gaattcgcggccgcttctagagGAAGTCTAACCGAAAGCAG</span></span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:8;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Bba_pos_r</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  class=GramE><span style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:
+  "Times New Roman";mso-bidi-font-family:"Times New Roman"'>actagtaAGTAGAGAGCGTTCACCG</span></span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:9;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Bba_seq_prefix_f</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Use <span class=SpellE>PBJ_seq_prefix_f</span><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:10;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Bba_seq_prefix_r</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>TGTTCTGCTGGTAGTGGTC<o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:11;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border-top:solid black 1.0pt;
+  border-left:solid black 1.0pt;border-bottom:none;border-right:none;
+  mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Bba_seq_suffix_f</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border-top:solid black 1.0pt;
+  border-left:none;border-bottom:none;border-right:solid black 1.0pt;
+  mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black .5pt;
+  padding:0in 5.4pt 0in 5.4pt;height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>Use <span class=SpellE>PBJ_seq_suffix_f</span><o:p></o:p></span></p>
+  </td>
+ </tr>
+ <tr style='mso-yfti-irow:12;mso-yfti-lastrow:yes;height:15.0pt'>
+  <td width=202 nowrap valign=bottom style='width:202.0pt;border:solid black 1.0pt;
+  border-right:none;mso-border-top-alt:solid black .5pt;mso-border-left-alt:
+  solid black .5pt;mso-border-bottom-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span class=SpellE><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'>Bba_seq_suffix_r</span></span><span
+  style='font-size:12.0pt;font-family:Calibri;mso-fareast-font-family:"Times New Roman";
+  mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
+  </td>
+  <td width=285 nowrap valign=bottom style='width:285.0pt;border:solid black 1.0pt;
+  border-left:none;mso-border-top-alt:solid black .5pt;mso-border-bottom-alt:
+  solid black .5pt;mso-border-right-alt:solid black .5pt;padding:0in 5.4pt 0in 5.4pt;
+  height:15.0pt'>
+  <p class=MsoNormal style='line-height:normal'><span style='font-size:12.0pt;
+  font-family:Calibri;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:
+  "Times New Roman"'>CACAAGCTGGAGTACAACTAC<o:p></o:p></span></p>
+  </td>
+ </tr>
+</table>
+</div>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'>&lt;<span class=GramE>data</span>
+table 5&gt;</p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal style='line-height:normal'><o:p>&nbsp;</o:p></p>
+<p class=MsoNormal><o:p>&nbsp;</o:p></p>
-<body class="c19"><p class="c3 c16"><span class="c2">Captains&rsquo; Log. iGEM 2014. Team NitroGENIUS. The Brauer Group.</span></p><p class="c3 c7 c16"><span class="c2"></span></p><p class="c3"><span class="c2">Day 1. 6/2/2014 (Monday) </span></p><ul class="c5 lst-kix_jsvobkvus7ai-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: design primers/plasmids </span></li><li class="c0"><span class="c1">Got done</span><span>: started primer/plasmid design</span></li></ul><p class="c3 c7"><span class="c15"></span></p><p class="c3"><span class="c2">Day 2. 6/3/2014 (Tuesday) </span></p><ul class="c5 lst-kix_bpa14sgapkx0-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Create and work on presentation</span></li><li class="c0"><span class="c1">Got done</span><<bospan>: Monsanto Presentation</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 3. 6/4/2014 (Wednesday)</span></p><ul class="c5 lst-kix_tmontj8so79x-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Monsanto Visit</span></li><li class="c0"><span class="c1">Got done</span><span>: Monsanto Visit</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 4. 6/5/014 (Thursday) </span></p><ul class="c5 lst-kix_vv1plefhvm43-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: More plasmid editing/primer design. Edit website. Plan out video series.</span></li><li class="c0"><span class="c1">Got done</span><span>: Finished pLight plasmid design and primers, sent to Cheryl for verification. Typed up brainstorming for video sessions, contacted WUTV about using equipment, contacted WashU Assembly Series director for permission to allow a Monsanto scientist to give a lecture during the Fall 2014 semester</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 5. 6/6/2014 (Friday) </span></p><ul class="c5 lst-kix_pg7viu9un1hn-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Get iGEM safety forms filled out. Go over primer/plasmid design. Order primers. Get keys/access to Brauer Hall. Design a timeline for the summer. </span></li><li class="c0"><span class="c1">Got done</span><span>: Finished design of plasmid PBJ001. Finished all (including sequencing) primer design. Take another look on Sunday, 6/8/2014.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 6. 6/9/2014 (Monday)</span></p><ul class="c5 lst-kix_n0j6o0qrtbio-0 start"><li class="c0"><span class="c4">Plan to do</span><span>:</span><span>&nbsp;Get iGEM safety forms filled out. Order primers. Get keys/access to Brauer Hall. Design a timeline for the summer. Complete lab safety forms for WashU. Design primers/plasmid map for positive/negative control. </span></li><li class="c0"><span class="c1">Got done</span><span>: Ordered primers (should arrive by Wednesday at the latest). Got keys/access to Brauer Hall. Completed lab safety forms for WashU. </span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 7. 6/10/2014 (Tuesday)</span></p><ul class="c5 lst-kix_t8wrn6z995vv-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Finish design of pos/neg control plasmids. Print Brauer lab safety form. Finalize proposal and send to Penn State iGEM contact along with project presentation. Design light experiment. </span></li><li class="c0"><span class="c1">Got done</span><span>: Designed primers for the positive and negative control plasmids. Printed safety form. Sent Penn State iGEM project info. PCR, ran visualization, and extracted pieces.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 8. 6/11/2014 (Wednesday)</span></p><ul class="c5 lst-kix_q3hqxbrf29cq-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Moon Lab Group meeting at 3PM. Work on finishing Gel DNA extraction, Dpn1, Purification, then One-Pot, then finally Transformation and plating (2 LB/cm plates). [</span><span class="c2">Realization! </span><span>We discovered today that in order for the ccaS/ccaR light system to function correctly, we need the PCC6803 genes </span><span class="c13">ho1 </span><span>and </span><span class="c13">pcyA</span><span>&nbsp;which are needed to heme into PCB (a chromophore) via a two step oxidation/reduction process. Ray is working on this aspect and we are planning to help/take over this part of his work. Our plan is to still transform plasmid PBJ and send it off for sequencing to make sure we have our designed plasmid. The next step would be to co-transform the two plasmids, one containing the </span><span class="c13">ho1 </span><span>and </span><span class="c13">pcyA</span><span>&nbsp;genes and the plasmid PBJ into </span><span class="c13">E. coli</span><span>&nbsp;and after sending off for sequencing, test the light promoter system. ]</span></li><li class="c0"><span class="c1">Got done</span><span>: Finished Gel DNA extraction and measured concentration using nanoquant tecan. Did not need to do Dpn1 nor purification. Put in one-pot after 12 noon. Transformed and incubated O/N. PCR for Ray&rsquo;s Biobrick plasmid for </span><span class="c13">ho1 </span><span>and </span><span class="c13">pcyA</span><span>&nbsp;parts. </span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 9. 6/12/2014 (Thursday)</span></p><ul class="c5 lst-kix_q3hqxbrf29cq-0"><li class="c0"><span class="c4">Plan to do</span><span>: Gel extract, one-pot for Ray&rsquo;s plasmid then transform and plate. pick PBJ002 colonies, start O/N liquid cultures. Work on negative and pos. control once primers arrive (check around noon). Also, grow a culture of Ray&rsquo;s BbaP DNA (AmpR) as well as Cheryl&rsquo;s pTet 100-T (where we get the TetR from)</span></li></ul><p class="c3"><span>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Plan out seq PCR for PBJ002.</span></p><ul class="c5 lst-kix_q3hqxbrf29cq-0"><li class="c0"><span class="c1">Got done</span><span>: Gel extract and one pot for Ray&rsquo;s plasmid. Re-ran PCR for Rays BbaP (SpeC_AraC_Bba) plasmid. Gel extract on new BbaP part. Did not yield bands- only saw a smear. Received primers for neg and pos controls- mixed appropriate primer concentrations. Cheryl started pTet liquid culture. Started BbaP liquid cultures. Electroporated original Bbap. Plated on 2 specR plates (100 uL) and (the rest). put into incubator. </span></li></ul><p class="c3"><span>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Planned out seq PCR for PBJ002. </span></p><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 10. 6/3/2014 (Friday)</span></p><ul class="c5 lst-kix_q3hqxbrf29cq-0"><li class="c0"><span class="c4">Plan to do</span><span>: meet at 9:15 in the ICARES to set up video conference with the Penn State. </span></li></ul><p class="c3 c11"><span>1</span><span class="c1">. PBJ002: Miniprep -&gt; PCR with sequencing primers before ICARES meeting</span></p><p class="c3 c7 c11"><span class="c1"></span></p><p class="c3 c10"><span class="c1">miniprep pTet and BbaP we made liquid cultures for yesterday. miniprep this along with the PBJ miniprep. </span></p><p class="c3 c11"><span>2. </span><span class="c1">pour gel for visual confirmation </span></p><p class="c3 c11"><span class="c1 c2">...ICARES meeting&hellip; then...</span></p><p class="c3 c11"><span>3. </span><span class="c1">take PCR product and run a gel (10 uL DNA, 2uL 6X dye) to check. 30 minutes.</span></p><p class="c3 c10"><span class="c1">Observe which PCR product in gel (which lane from tube) is of the right length.</span></p><p class="c3 c10"><span class="c9">Purify PCR product.</span></p><p class="c3 c11"><span class="c1 c2">Send miniprepped plasmid &nbsp;</span><span class="c1 c2 c9">AND purified PCR</span><span class="c1 c2">&nbsp;off for sequencing of PBJ002. </span></p><p class="c3 c11"><span>4. work on both the Positive and negative controls.</span></p><p class="c3 c11"><span>5.</span><span class="c1">&nbsp;Pick colonies and start O/N culture for Spec_AraC_Bba. </span></p><ul class="c5 lst-kix_q3hqxbrf29cq-0"><li class="c0"><span class="c1">Got done</span><span>: From the above mentioned steps: Steps 1- 5. </span></li></ul><ul class="c5 lst-kix_q3hqxbrf29cq-1 start"><li class="c3 c6"><span>For the Pos and Neg controls, we did PCR -&gt; nanoquant measure after purification step (reference pos/neg control step documents) </span><span class="c4">TetR backbone not present. Re-run tomorrow. </span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 11. 6/14/2014 (Saturday)</span></p><ul class="c5 lst-kix_9s5e088b0z9v-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: </span></li></ul><ul class="c5 lst-kix_9s5e088b0z9v-1 start"><li class="c3 c6"><span>Miniprep -&gt; Visualize seq PCR product of Spec_AraC_Bba</span></li></ul><ul class="c5 lst-kix_9s5e088b0z9v-2 start"><li class="c3 c8"><span>could not run seq PCR on this because seq primers do not work. Re-design sequencing primers and order them.</span></li></ul><ul class="c5 lst-kix_9s5e088b0z9v-1"><li class="c3 c6"><span class="c4">Re-run PCR for TetR backbone. Failed yesterday. </span></li><li class="c3 c6"><span class="c4 c9">One pot Pos control</span></li><li class="c3 c6"><span>One pot Neg control</span></li></ul><p class="c3 c11"><span>...Later...</span></p><ul class="c5 lst-kix_9s5e088b0z9v-1"><li class="c3 c6"><span>Transform/plate neg control</span></li><li class="c3 c6"><span class="c9">Transform/plate pos control</span></li></ul><ul class="c5 lst-kix_9s5e088b0z9v-0"><li class="c0"><span class="c1">Got done</span><span>: </span></li></ul><ul class="c5 lst-kix_9s5e088b0z9v-1 start"><li class="c3 c6"><span>Miniprepped and measured concentration on SAB product. </span></li></ul><ul class="c5 lst-kix_9s5e088b0z9v-2 start"><li class="c3 c8"><span class="c17">Project SAB is on hold until primers arrive.</span></li></ul><ul class="c5 lst-kix_9s5e088b0z9v-1"><li class="c3 c6"><span>Re ran PCR on the TetR backbone. </span></li><li class="c3 c6"><span>sent new redesigned sequencing primers for SAB to Ray for ordering.</span></li><li class="c3 c6"><span>Gel Extraction/Recovery on TetR backbone PCR product </span></li><li class="c3 c6"><span>Dpn1 for TetR backbone </span></li><li class="c3 c6"><span>TetR purification and measurement= 4.4 ng/uL </span></li></ul><ul class="c5 lst-kix_9s5e088b0z9v-2 start"><li class="c3 c8"><span class="c17">Project Positive Control is on hold until we can re-evaluate the primers and find the error/form a new plan of approach.</span></li><li class="c3 c8"><span class="c18">Correction: re-did PCR for TetR backbone again. (was the reason we couldn&rsquo;t get it to work before because we used the wrong plasmid?)</span></li></ul><ul class="c5 lst-kix_9s5e088b0z9v-3 start"><li class="c3 c20"><span class="c18">gel only yielded bands indicating primers.</span></li></ul><ul class="c5 lst-kix_9s5e088b0z9v-1"><li class="c3 c6"><span class="c1">One pot Negative control (-) lacZ</span></li><li class="c3 c6"><span class="c1">Transform/plated Negative control (-) lacZ</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 12. 6/15/2014 (Sunday)</span></p><ul class="c5 lst-kix_qsjdqtsjwd5p-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: </span></li></ul><ul class="c5 lst-kix_qsjdqtsjwd5p-1 start"><li class="c3 c6"><span>TetR backbone for positive control run from PCR- &gt;Transformation/Plating</span></li><li class="c3 c6"><span>Pick Neg Control colonies. </span></li></ul><ul class="c5 lst-kix_qsjdqtsjwd5p-2 start"><li class="c3 c8"><span>Start liquid cultures for O/N </span></li></ul><ul class="c5 lst-kix_qsjdqtsjwd5p-0"><li class="c0"><span class="c1">Got done</span><span>:</span></li></ul><ul class="c5 lst-kix_qsjdqtsjwd5p-1 start"><li class="c3 c6"><span>(+) TetR backbone from TetR-100T miniprepped plasmid: PCR -&gt; DNA purification</span></li><li class="c3 c6"><span>Picked Neg Control colonies. </span></li></ul><ul class="c5 lst-kix_qsjdqtsjwd5p-2 start"><li class="c3 c8"><span>Started liquid cultures for O/N </span></li></ul><p class="c3"><span class="c2">&nbsp;</span></p><p class="c3"><span class="c2">Day 13. 6/16/2014 (Monday)</span></p><ul class="c5 lst-kix_a0ubbid7t2g3-0 start"><li class="c0"><span class="c4">Plan to do</span><span>:</span></li></ul><ul class="c5 lst-kix_a0ubbid7t2g3-1 start"><li class="c3 c6"><span>Negative control: freeze and miniprep -&gt; Send to Medical School for sequencing</span></li><li class="c3 c6"><span>Positive control: One-pot -&gt; Transform/plate</span></li></ul><ul class="c5 lst-kix_a0ubbid7t2g3-0"><li class="c0"><span class="c1">Got done</span><span>:</span></li></ul><ul class="c5 lst-kix_a0ubbid7t2g3-1 start"><li class="c3 c6"><span>Freeze and miniprep negative control</span></li><li class="c3 c6"><span class="c2">Negative Control</span><span>: Ran Sequencing PCR for (-) lacZ</span></li></ul><ul class="c5 lst-kix_a0ubbid7t2g3-2 start"><li class="c3 c8"><span>Visualization gel (smear)</span></li><li class="c3 c8"><span>Rerun sequencing PCR with new settings- failure</span></li></ul><p class="c3 c10"><span>Redux starting with DPN1 (-) lacZ #5 </span></p><ul class="c5 lst-kix_a0ubbid7t2g3-2"><li class="c3 c8"><span>Purify</span></li><li class="c3 c8"><span>Nanoquant</span></li><li class="c3 c8"><span>Blunt end ligation</span></li><li class="c3 c8"><span class="c1">Transformation/Electroporation and plating on </span><span class="c1 c2">cm</span><span class="c1">&nbsp;plates </span></li></ul><ul class="c5 lst-kix_a0ubbid7t2g3-1"><li class="c3 c6"><span class="c2">Positive Control </span><span>(EYFP + TetR backbone) Run One Pot</span></li></ul><ul class="c5 lst-kix_a0ubbid7t2g3-2 start"><li class="c3 c8"><span>Transformed</span></li><li class="c3 c8"><span>plated 100uL, rest at 8:05pm</span></li><li class="c3 c8"><span>Incubate o/n</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 14. 6/17/2014 (Tuesday)</span></p><ul class="c5 lst-kix_atwdcoxr33pn-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Pick both Positive and Negative Control colonies</span></li></ul><ul class="c5 lst-kix_atwdcoxr33pn-1 start"><li class="c3 c6"><span>Positive: </span><span class="c2">kan antibiotic</span></li><li class="c3 c6"><span>Negative: </span><span class="c2">cm antibiotic</span></li></ul><ul class="c5 lst-kix_atwdcoxr33pn-2 start"><li class="c3 c8"><span class="c4 c2">no colonies seen as of 10 AM</span></li><li class="c3 c8"><span class="c4 c2">colonies seen at 12 noon! </span></li></ul><ul class="c5 lst-kix_atwdcoxr33pn-0"><li class="c0"><span class="c1">Got done</span><span>: </span></li></ul><ul class="c5 lst-kix_atwdcoxr33pn-1 start"><li class="c3 c6"><span>redesigned new PBJ003 plasmid map and designed/verified new sequencing primers. Sent them to Cheryl for verification. SAB primers have not yet arrived. </span><span>Liquid cultures into the incubator by 9PM</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 15. 6/18/14 (Wednesday) -Worldly Wednesday!</span></p><ul class="c5 lst-kix_kegcie22s4uf-0 start"><li class="c0"><span class="c4">Plan to do</span><span>:</span></li></ul><ul class="c5 lst-kix_kegcie22s4uf-1 start"><li class="c3 c6"><span>Get both Pos and Neg frozen and miniprepped -&gt; send off for sequencing</span></li></ul><ul class="c5 lst-kix_kegcie22s4uf-2 start"><li class="c3 c8"><span>concurrently with SAB?</span></li></ul><ul class="c5 lst-kix_kegcie22s4uf-1"><li class="c3 c6"><span>Go to Worldy Wednesday Lunch hosted by Andrew Ng </span></li><li class="c3 c6"><span>Work on SAB?</span></li></ul><ul class="c5 lst-kix_kegcie22s4uf-0"><li class="c0"><span class="c1">Got done</span><span>: </span></li></ul><ul class="c5 lst-kix_kegcie22s4uf-1 start"><li class="c3 c6"><span>Froze both Pos and Neg and miniprepped. </span></li></ul><ul class="c5 lst-kix_kegcie22s4uf-2 start"><li class="c3 c8"><span>Pos process went to sequencing. </span></li><li class="c3 c8"><span>Negative control: start at seq PCR. </span></li></ul><ul class="c5 lst-kix_kegcie22s4uf-1"><li class="c3 c6"><span>Ran SAB SeqPCR. </span></li></ul><ul class="c5 lst-kix_kegcie22s4uf-2 start"><li class="c3 c8"><span>negative results</span></li></ul><ul class="c5 lst-kix_kegcie22s4uf-1"><li class="c3 c12 c6"><span>Benjamin Colony PCR negative control (- lacZ) from old plates</span></li></ul><p class="c3 c12 c7"><span></span></p><p class="c3 c12"><span class="c2">Day 16. 6/19/14 (Thursday)</span></p><ul class="c5 lst-kix_5ru64txmw2lz-0 start"><li class="c0 c12"><span class="c4">Plan to do</span><span>: </span></li><li class="c0 c12"><span class="c1">Got done</span><span>:</span></li></ul><ul class="c5 lst-kix_5ru64txmw2lz-1 start"><li class="c3 c12 c6"><span>Re-designed the SEQ primers for SAB. </span></li><li class="c3 c12 c6"><span>CPCR for (-)lacZ</span></li></ul><ul class="c5 lst-kix_5ru64txmw2lz-2 start"><li class="c3 c12 c8"><span>move on to (-)cph8</span></li></ul><ul class="c5 lst-kix_5ru64txmw2lz-1"><li class="c3 c12 c6"><span>begin work on PBJ003</span></li></ul><p class="c3 c12 c7"><span></span></p><p class="c3 c12"><span class="c2">Day 17. 6/20/14 (Friday)</span></p><ul class="c5 lst-kix_5ru64txmw2lz-0"><li class="c0"><span class="c4">Plan to do</span><span>: </span></li><li class="c0"><span class="c1">Got done</span><span>:</span></li></ul><ul class="c5 lst-kix_5ru64txmw2lz-1 start"><li class="c3 c6"><span>PCR and gel extracted correct EYFP</span></li><li class="c3 c6"><span>working on getting piece Ori PCR -&gt; gel extract and setting up one-pot for PBJ003</span></li><li class="c3 c6"><span>(-) cph8 took all the way through DNA clean &amp; concentrate and added wrong wash buffer</span></li></ul><ul class="c5 lst-kix_5ru64txmw2lz-2 start"><li class="c3 c8"><span>5&micro;l purified DNA remaining in -20 (inside cph8 box)</span></li></ul><p class="c3 c7 c10"><span></span></p><p class="c3"><span class="c2">Day 18. 6/21/14 (Saturday)</span></p><ul class="c5 lst-kix_jplp7lygncqy-0 start"><li class="c0"><span class="c4">Plan to do</span><span>:</span></li><li class="c0"><span class="c1">Got done</span><span>:</span></li></ul><ul class="c5 lst-kix_jplp7lygncqy-1 start"><li class="c3 c6"><span>Negative Control:DPN1, purify, nanoquant measure, blunt end ligation (BEL) and transform/plate on </span><span class="c2">cm</span><span>&nbsp;</span></li><li class="c3 c6"><span>PBJ003: PCR -&gt; DNA Recover and Nanoquant measure. Digestion/Ligation (One-Pot) all pieces. Transformation/Plating on </span><span class="c2">cm</span></li></ul><p class="c3 c7"><span class="c2"></span></p><p class="c3"><span class="c2">Day 19. 6/22/14 (Sunday)</span></p><ul class="c5 lst-kix_dlh3l7v040tq-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Pick colonies start liquid colonies for -cph8, pbj003</span></li><li class="c0"><span class="c1">Got done</span><span>: Pick colonies start liquid colonies for -cph8, pbj003</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 20. 6/23/14 (Monday)</span></p><ul class="c5 lst-kix_5k3if7w24bx9-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Freeze, miniprep, sequencing pcr, visualization gel, send for sequencing</span></li><li class="c0"><span class="c1">Got done</span><span>: negative control, pbj003: Freeze, miniprep, sequencing pcr, visualization gel, send for sequencing</span></li></ul><ul class="c5 lst-kix_5k3if7w24bx9-1 start"><li class="c3 c6"><span>SAB: Sequencing PCR, visualization gel</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 21. 6/24/14 (Tuesday)</span></p><ul class="c5 lst-kix_tnagh66xnbun-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: design experimental methods and procedures, design around hybrid promoter, find lacI resistant strain</span></li><li class="c0"><span class="c1">Got done</span><span>: </span></li></ul><ul class="c5 lst-kix_tnagh66xnbun-1 start"><li class="c3 c6"><span>did modified SEQPCR on SAB and ran visualization gel- confirmed S and B present, A not present</span></li><li class="c3 c6"><span>Ran PCR on part AraC/p15a ( part A), extracted and DNA recovered along with nanoquant measurements. </span></li><li class="c3 c6"><span class="c1">S,A,B Digestion</span></li><li class="c3 c6"><span class="c1">S+A+B Ligation into </span><span class="c1 c2">SAB2</span></li><li class="c3 c6"><span class="c18">Transformation and Plating of </span><span class="c2 c18">SAB2 (failed)</span></li></ul><p class="c3 c7"><span class="c2 c18"></span></p><p class="c3"><span class="c2">Day 22. 6/25/14 (Worldly Wednesday)</span></p><ul class="c5 lst-kix_vsah3hmcuwyk-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Transform SAB2 C and D. Check sequencing results for the negative control and PBJ003 if they come in today. </span></li><li class="c0"><span class="c1">Got done</span><span>: Verfied sequences for some positive control #4, some PBJ003. Sent T2 and E4 of PBJ003 in for sequencing confirmation, send in (-) lacZ sequencing on (-)cph8 plasmid, transformed and plated SAB2 C and D. Design primers for hybrid promoter</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 23. 6/26/14 (Thursday)</span></p><ul class="c5 lst-kix_yffclurgw5vk-0 start"><li class="c0"><span class="c4">Plan to do:</span><span>&nbsp;Pick and grow colonies for liquid culture</span></li><li class="c0"><span class="c1">Got done: </span><span>Grew liquid Colonies for SAB2. Work on website. Look up experimental procedures</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 24. 6/27/14 (Friday)</span></p><ul class="c5 lst-kix_yffclurgw5vk-0"><li class="c0"><span class="c4">Plan to do:</span><span>&nbsp;Freeze and miniprep SAB2; Sequencing PCR, Send for sequencing</span></li><li class="c0"><span class="c1">Got done: </span><span>Freeze and miniprep SAB2; Sequencing PCR, </span><span class="c4">PCR fail, will re run with a gradient. </span><span>Verified completely PBJ003 #2, #4. (-) lacZ funky. Received primers for hybrid promoter</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 25. 6/28/14 ( Saturday)</span></p><ul class="c5 lst-kix_yffclurgw5vk-0"><li class="c0"><span class="c4">Plan to do:</span><span>&nbsp;Run Sequencing PCR (gradient) for SAB2C #2. Send out purified PCR product for Sequencing. Set up experimental procedures, write out experiment protocol.</span></li><li class="c0"><span class="c1">Got done: </span><span>Ran sequencing PCR with a gradient for SAB2 C#2. Verified negative control completely finished. </span></li></ul><p class="c3 c7"><span class="c1"></span></p><p class="c3"><span class="c2">Day 26. 6/29 /14 ( Sunday)</span></p><ul class="c5 lst-kix_yffclurgw5vk-0"><li class="c0"><span class="c4">Plan to do:</span><span>&nbsp;Run SEQPCR (gradient) for SAB2C #2 and visualize. </span></li><li class="c0"><span class="c1">Got done: </span><span>Ran SEQPCR for SAB2C #1. Same results on from 6/28/14. Redesigned primers for new plasmid (not using SAB anymore) called pHOP</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 27. 6/30 /14 ( Monday)</span></p><ul class="c5 lst-kix_yffclurgw5vk-0"><li class="c0"><span class="c4">Plan to do:</span><span>&nbsp;</span></li><li class="c0"><span class="c1">Got done: </span><span>picked ice mix from Ray&rsquo;s pSB1C3-BBa_M30109 frozen stock. Preparing for miniprep tomorrow morning (0800). </span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 28. 7/1 /14 ( Tuesday)</span></p><ul class="c5 lst-kix_yffclurgw5vk-0"><li class="c0"><span class="c4">Plan to do:</span><span>&nbsp;Miniprep pSB1C3-BBa_M30109. Do a digestion and gel visualization step for pSB1C3-BBa_M30109 to see if its legit. Start PCR and steps for PBJ_Hybrid (PBJ003 + hybrid promoter)</span></li><li class="c0"><span class="c1">Got done: </span><span>Saw nothing in the first digestion, could not re-run digestion due to no enzyme left. PCR, DPN1, Clean, BEL of PBJ_Hyb_001, transform, plate</span></li></ul><p class="c3 c7"><span class="c1 c2"></span></p><p class="c3"><span class="c2">Day 29. 7/2/14 (Worldly Wednesday)</span></p><ul class="c5 lst-kix_l5wvx3c462ts-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: make LB/cm plates (follow protocol), look at plated cultures and wrap when necessary. pHOP primers should arrive today so we can get started working on that</span></li><li class="c0"><span class="c1">Got done</span><span>: Pulled plates (2) from incubator, wrapped them and put into 4 fridge. Picked colonies for PBJ_hyb_001. started pHOP procedures. </span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 30. 7/3/14 (Thursday)</span></p><ul class="c5 lst-kix_x1igvv7x84s6-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Miniprep -&gt; send off for sequencing for PBJ_hyb_001. One pot then Transform and plate pHOP</span></li><li class="c0"><span class="c1">Got done</span><span>: Miniprep-&gt; visualize for PBJ_hyb_001. Plan on preparing for sequencing on Sunday. One pot, transformed and plated pHOP. </span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 31. 7/4/14 (Friday. &lsquo;Murica.)</span></p><ul class="c5 lst-kix_zihm5tvntbif-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Pick pHOP colonies and start liquid cultures with kan for O/N.</span></li><li class="c0"><span class="c1">Got done</span><span>: Start liquid cultures</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 32. 7/5/14 (Saturday)</span></p><ul class="c5 lst-kix_zf88vxbhzn7x-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Freeze -&gt; Run SEQPCR Gel on pHOP.</span></li><li class="c0"><span class="c1">Got done</span><span>: pHOP gel bands are of the wrong length. </span></li></ul><p class="c3 c7"><span class="c2"></span></p><p class="c3"><span class="c2">Day 33. 7/6/14 (Sunday)</span></p><ul class="c5 lst-kix_cc5xl3s45cjm-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Get sequencing ready for PBJ_hyb_001, re run pHOP SEQPCR</span></li><li class="c0"><span class="c1">Got done</span><span>: Go sequencing ready for hybrid promoter. pHOP SEQPCR gel visualization not of correct length. </span></li></ul><p class="c3 c7"><span class="c2"></span></p><p class="c3"><span class="c2">Day 34. 7/7/14 (Monday)</span></p><ul class="c5 lst-kix_cc5xl3s45cjm-0"><li class="c0"><span class="c4">Plan to do</span><span>: One Pot enhanced pHOP2/ Colony PCR pHOP3</span></li><li class="c0"><span class="c1">Got done</span><span>: Colony PCR for pHOP3 showed wrong bands. Enhanced One Pot for pHOP2, transformed, plated on LB/Kan</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 35. 7/8/14 (Tuesday)</span></p><ul class="c5 lst-kix_cc5xl3s45cjm-0"><li class="c0"><span class="c4">Plan to do</span><span>: Start Liquid culture for pHOP2</span></li><li class="c0"><span class="c1">Got done</span><span>: Started Liquid culture for pHOP2, Colony PCR pHOP2, Verified Sequencing for PBJ_hyb_001 3F, ready to go.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 36. 7/9/14 (Wednesday)</span></p><ul class="c5 lst-kix_cc5xl3s45cjm-0"><li class="c0"><span class="c4">Plan to do</span><span>: Sequencing pcr for pHOP2, send for sequencing</span></li><li class="c0"><span class="c1">Got done</span><span>: Sequencing PCR failed, do a modified digest of BBa_M30109 using BSG1, failed. Work on degradation tag design (finished)</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 37. 7/10/14 (Thursday)</span></p><ul class="c5 lst-kix_6s2krrh2clpb-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Get a protocol written for testing of PBJ_hyb_001 (light and dark)</span></li><li class="c0"><span class="c1">Got done</span><span>: Protocol for testing written out. Started liquid cultures in light/dark</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 38. 7/11/14 (Friday)</span></p><ul class="c5 lst-kix_8udtkhf2dbar-0 start"><li class="c0"><span class="c4">Plan to do</span><span>:</span></li><li class="c0"><span class="c1">Got done</span><span>: Tested hybrid and PBJ003 without chromophore. designed primers for PBJ004 (with Ptet promoter Ptet-pp+ plasmid from Dr. Moon to control EYFP). </span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 39. 7/14/14 (Monday)</span></p><ul class="c5 lst-kix_9atxlh6cz8gg-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Begin cloning for degradation tags on PBJ003 AND PBJ_hyb_001</span></li><li class="c0"><span class="c1">Got done</span><span>: Started PCR for Degradation Tags as well as Ptet-pp (same as: Ptet-T*2) minus the TetR (as a control). Designed light-box</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 40. 7/15/14 (Tuesday)</span></p><ul class="c5 lst-kix_6k3a397ugjom-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: meet with Andrew to go over light-box ordering. Continue cloning projects. </span></li><li class="c0"><span class="c1">Got done</span><span>: cloning project up to Purification after Dpn1- no DNA present so re-did from PCR. </span></li></ul><p class="c3"><span class="c2">Day 41. 7/16/14 (Wednesday)</span></p><ul class="c5 lst-kix_iznc9vnsc2z-0 start"><li class="c0"><span>Plan to do: Keep cloning</span></li><li class="c0"><span>Got done: kept working on pet(-)tetR, PBJ003_DegTetR, and PB_hyb_001 DegTetR up to transformation and plating</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 42. 7/17/14 (Thursday)</span></p><ul class="c5 lst-kix_1y65tr9fq1c6-0 start"><li class="c0"><span class="c4">Plan to do</span><span>: Check on and wrap plates. Start liquid cultures for both degradation tags, ptet(-)tetR and biobrick part from shipped frozen stock.</span></li><li class="c0"><span class="c1">Got done</span><span>: Checked plates, wrapped them around 10:20 AM. Started Liquid culture at 6pm.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 43. 7/18/14 (Friday)</span></p><ul class="c5 lst-kix_1y65tr9fq1c6-0"><li class="c0"><span class="c4">Plan to do</span><span>: Miniprep, sequencing PCR degradation tags, ptet(-). Miniprep Biobrick part, digestion &amp; ligation alongside PFM109 &amp; transform</span></li><li class="c0"><span class="c1">Got done</span><span>: Sequencing PCR for degradation tags. ptet(-) doesnt look right. Only miniprepped biobrick part.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 44. 7/19/14 (Saturday)</span></p><ul class="c5 lst-kix_1y65tr9fq1c6-0"><li class="c0"><span class="c4">Plan to do</span><span>: Sequencing grad PCR for ptet(-), Sequential Digestion for pFM109, Bba_K1017726 -&gt; Gel Extract -&gt; Ligation -&gt; Transform &amp; plate</span></li><li class="c0"><span class="c1">Got done</span><span>: Digested with EcoR1, Digested with SpeI, Ligation; Sequencing PCR for ptet(-): piece size not as expected ran colony pcr- band length still indicates tetR present</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 45. 7/20/14 (Sunday)</span></p><ul class="c5 lst-kix_k39z8c0z9nv-0 start"><li class="c0"><span>Plan to do: Prep DegTag products PBJ003 and hybrid for Sequencing. Transform and Plate pChrom.</span></li><li class="c0"><span>Got done: Degradation Tag on TetR for both PBJ003 and hybrid are ready for sequencing. Transform and plate pChrom on lb/kanR plates. Put in 37 at 7:50pm</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 46. 7/21/14 (Monday)</span></p><ul class="c5 lst-kix_eky4cy64bu8k-0 start"><li class="c0"><span>Plan to do: send tags off for sequencing. Start ptet(-) tetR cloning steps. Start liquid cultures for Chromophore project. </span></li><li class="c0"><span>Got done: tags sent for sequencing. finished from PCR to dpn1 on ptet (-) tetR. colony pcr for chromophore failed. Redo ligation and transformation</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 47. 7/22/14 (Tuesday)</span></p><ul class="c5 lst-kix_1wkl9z7rug8w-0 start"><li class="c0"><span>Plan to do: redo colony pcr for chromophore. Work on ptet(-)tetR positive control.</span></li><li class="c0"><span>Got done: kept working on ptet(-)tetR positive control- dpn1 and purify,transformed and plated. Not enough colonies for chromophore to colony pcr. Started liquid culture at night.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 48. 7/23/14 (Wednesday)</span></p><ul class="c5 lst-kix_yuqzvf2rvpom-0 start"><li class="c0"><span>Plan to do: Miniprep and sequencing pcr for chromophore. Check and wrap plates, start liquid cultures around 8 or 9. </span></li><li class="c0"><span>Got done: Miniprep and Sequencing PCR for chromophore. Prepped for sequencingstarted (-)tetR liquid cultures. worked on PCR seq for Degradation tags on PBJ003 and Phybrid. &nbsp;&ldquo;need&rdquo; to purify before sending pcr product. </span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 49. 7/24/14 (Thursday)</span></p><ul class="c5 lst-kix_yuqzvf2rvpom-0"><li class="c0"><span>Plan to do: Purify PCR products. Miniprep and check sequencing pcr for (-)tetR. Send all for sequencing</span></li><li class="c0"><span>Got done: Purified PCR product for chrom and degradation tags. Prepped for sequencing. Miniprep and run sequencing PCR for (-)tetR (no bands)</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 50. 7/25/14 (Friday)</span></p><ul class="c5 lst-kix_1en1b0ghue45-0 start"><li class="c0"><span>Plan to do: Gradient SEQPCR on (-)tetR, run a gel visualizatoin, prep for sequencing.</span></li><li class="c0"><span>Got done: Ran gradient on (-)tetR #3 and 4 with gel visualization, saw bands at wrong length. doing colony PCR tomorrow. </span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 51. 7/26/14 (Saturday)</span></p><ul class="c5 lst-kix_esnh4vdjimna-0 start"><li class="c0"><span>Plan to do: Brute Force Colony PCR (basically picking 32 colonies!). Freeze and Miniprep correct ones. </span></li><li class="c0"><span>Got done: Brute Force Colony PCR and gel visualization finished.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 52. 7/27/14 (Sunday)</span></p><ul class="c5 lst-kix_6cjq62pzygtv-0 start"><li class="c0"><span>Plan to do: Start all cloning procedures by taking #4 of the degradation tags from hybrid and pBJ003. &nbsp;Prep for sequencing the miniprepped &nbsp;(-)tetR DNA samples (#9,12,15,16). </span></li><li class="c0"><span>Got done: Prep (-)tetR for sequencing. Start cloning PBJ004 and PBJ_hyb_002</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 53. 7/28/14 (Monday)</span></p><ul class="c5 lst-kix_37xjlil3nv1-0 start"><li class="c0"><span>Plan to do: Digest Biobrick part to check for correct lengths&hellip; continue cloning of PBJ004 and PBJ_hyb_002 with the new ptet promoter insertion. Look more at LED grid coding. </span></li><li class="c0"><span>Got done: Digestion of biobrick part only showed a smear. Still send both miniprep and pcr product for sequencing </span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 54. 7/29/14 (Tuesday)</span></p><ul class="c5 lst-kix_h2or0sv4eu23-0 start"><li class="c0"><span>Plan to do: Check plates in the morning, wrap. Start cloning for PBJ005 and Hybrid 003. Start liquid cultures at night.</span></li><li class="c0"><span>Got done: plates checked and wrapped. Sanity check #2 for biobrick part, doing a double digestion. Started cloning of PBJ005 and Hybrid003. Liquid cultures started. Retransform003, order new sequencing primer, redo double digestion.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 55. 7/30/14 (Wednesday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0 start"><li class="c0"><span>Plan to do: Freeze, miniprep, send off for sequencing of PBJ004 and Hybrid002. Delete the CAC in the maps for PBJ003 with DegTagTetR and PBJ_hyb with DegTagTetR. Get iGEM calendar set up. Email Pakrasi and Moon about setting up final iGEM meeting</span></li><li class="c0"><span>Got done: Start Modeling of light promoter system. (-) tetR still no luck. &nbsp;Sequence verified pCHROM #1. Freeze miniprep, PBJ004, Hybrid 002. Start liquid cultures for PBJ005, Hybrid 003.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 56. 7/31/14 (Thursday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Make LB/ (spec + cm) &amp; (spec + kan) plates. Seq PCR for (-) tetR. Freeze miniprep PBJ005, Hybrid003. Send PBJ004, PBJ005, Hybrid 003, Hybrid 002, (-) tetR for sequencing. Design experimental protocol. Transform.</span></li><li class="c0"><span>Got done: Made plates. Miniprep PBJ005, hybrid 003. Sequencing PCR, gel visualization for PBJ005, hybrid003, (-) tetR. Gel visualize. Gradient PCR hybrid003, (-)tetR, visualize. Found Hybrid003 1E, 1F, (-)tetR wrong bands. Prep PBJ004, PBJ005, hybrid002 for sequencing. Transform/ plate pchrom + (pos/neg/pbj003,phybrid).</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 57. 8/1/14 (Friday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Prep Hybrid003 for sequencing. Send all in before 9. Pick cotransformed colonies. Start liquid culture.</span></li><li class="c0"><span>Got done: Sent for sequencing. Colonies not growing well. Colony PCR pchrom3 (from 7/29/14). Work on modeling.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 58. 8/2/14 (Saturday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Check plates. Wrap. Start liquid cultures</span></li><li class="c0"><span>Got done: Wrap colony PCR plate. Transformants not growing. *** Realization, pCHROM has kan resistance, so using the wrong plates the whole time. Started liquid cultures for pCHROM3, pCHROM1.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 59. 8/3/14 (Sunday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Miniprep pCHROM3 and 1</span></li><li class="c0"><span>Got done: Freeze pCHROM3, Miniprep pCHROM3 and 1. Sequencing results for new Ptet bad</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 60. 8/4/14 (Monday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Make Kan/CM plates. Check Sequence for pCHROM3. Transform and plate for experiment</span></li><li class="c0"><span>Got done: Made plates. Sequence PCR #1 showed multiple bands. Sequence PCR #2 showed wrong bands. Started new Ptet stuff over again from PCR. Transformed and plate experimental stuff</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 61. 8/5/14 (Tuesday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Wrap Plates. Miniprep PBJ003, PBJ_Hyb_001. Redo cloning for new Ptet plasmids. Start liquid cultures for experimentation. Work on presentation</span></li><li class="c0"><span>Got done: Wrap plates. Miniprep. Redo PBJ005</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 62. 8/6/14 (Wednesday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Run experiment. Induce. Check data</span></li><li class="c0"><span>Got done: Lighted growth chamber in use at 30 degrees. Cheryl advise to abort util tomorrow&hellip; Work on presentation. Purify, Transform PBJ005 redo. Design PBJpos</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 63. 8/7/14 (Thursday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Run experiment. Induce. Check data</span></li><li class="c0"><span>Got done: Ran experiment. Dilute to OD .1, grow for 2 hours, induction. Start PBJ005 liquid cultures, measure fluorescence. NO FLUORESCENCE</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 64. 8/8/14 (Friday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Send PBJ005 for sequencing</span></li><li class="c0"><span>Got done: Froze, miniprep, Sequencing PCR. Send Purified Seq PCR product for sequencing. Jeffrey&rsquo;s last day</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 65. 8/11/14 (Monday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Start cloning for PBJpos</span></li><li class="c0"><span>Got done: PCR, visualize, DPN1, purify, Blunt end ligate. Transform PBJpos. Also Transform PSL2264</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 66. 8/12/14 (Tuesday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Start liquid culture for PBJpos, PSL2264</span></li><li class="c0"><span>Got done: Wrapped plates. Checked sequence results for PBJ005=bad. Started liquid cultures for PBJpos</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 67. 8/13/14 (Wednesday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Miniprep, send for sequencing, start liquid cultures for experimentation</span></li><li class="c0"><span>Got done: Miniprepped and sent PBJpos #1-#4 for sequencing. Started liquid cultures for pSL2264, PBJpos 1-4, current positive control and negative control for experimentation tomorrow.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 68. 8/14/14 (Thursday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Run fluorescence tests for old and new positive controls. Sanity check for pSL2264</span></li><li class="c0"><span>Got done: pSL2264 not fluorescing as expected. PBJpos slightly better, Old positive control and negative control as expected. Induce pSL2264 with IPTG. Design new PBJ003 with deletion of competing TSS</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 69. 8/15/14 (Friday)</span></p><ul class="c5 lst-kix_woibo694d9q9-0"><li class="c0"><span>Plan to do: Design primers for new PBJ plasmids with modified RBS. Plan out fall semester work schedule.</span></li><li class="c0"><span>Got done: Design primers for new PBJ plasmids with modified RBS.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 70. 8/25/14 (Monday)</span></p><ul class="c5 lst-kix_g0w1w45ynsy6-0 start"><li class="c0"><span>Plan to do: Start cloning for PBJposrbs (new positive control with enhanced RBS) until transformation</span></li><li class="c0"><span>Got done: PCR, gel visualize, DPN1, Ligation, Transformation, Plating. Work on Protocol section for website.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 71. 8/26/14 (Tuesday)</span></p><ul class="c5 lst-kix_g0w1w45ynsy6-0"><li class="c0"><span>Plan to do: Start Liquid Culture</span></li><li class="c0"><span>Got done: Start liquid culture. Work on protocols.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 72. 8/27/14 (Wednesday)</span></p><ul class="c5 lst-kix_g0w1w45ynsy6-0"><li class="c0"><span>Plan to do: Miniprep. Run sequencing PCR, send for sequencing</span></li><li class="c0"><span>Got done: Miniprep. Run sequencing PCR, results bad. Rerun sequencing PCR with correct primers.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 73. 8/28/14 (Thursday)</span></p><ul class="c5 lst-kix_g0w1w45ynsy6-0"><li class="c0"><span>Plan to do: Visualize sequencing PCR. PCR purify, prep for sequencing. Send</span></li><li class="c0"><span>Got done: visualize sequencing PCR. PCR purify, prep for sequencing. Work on title and abstract</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 74. 8/29/14 (Friday)</span></p><ul class="c5 lst-kix_g0w1w45ynsy6-0"><li class="c0"><span>Plan to do: Send in for sequencing (morning). Make electrocompetent cells</span></li><li class="c0"><span>Got done: Send in for sequencing. Make electrocompetent cells. Work on title and abstract.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 75. 9/1/14 (Monday)</span></p><ul class="c5 lst-kix_g0w1w45ynsy6-0"><li class="c0"><span>Plan to do: Sequence verify PBJpos_RBS. Start liquid cultures for experimentation.</span></li><li class="c0"><span>Got done: Sequence verify PBJpos_RBS. Start liquid cultures for experimentation. Final safety forms due.!</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 76. 9/2/14 (Tuesday)</span></p><ul class="c5 lst-kix_g0w1w45ynsy6-0"><li class="c0"><span>Plan to do: Test for Fluorescence of newest positive control. </span></li><li class="c0"><span>Got done: tested for fluorescence. Dilute to 0.1 grew an extra 6 hours, tested again. EYFP did not fluoresce as expected. Compard 5&rsquo; UTRs between different positive controls. redesign primers in order to change pTet from tabor to moon&rsquo;s.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 77. 9/4/14 (Thursday)</span></p><ul class="c5 lst-kix_faphe7ovfpq5-0 start"><li class="c0"><span>Plan to do: Start cloning once primer&rsquo;s are received.</span></li><li class="c0"><span>Got done: Organize primers for assembly. Start cloning for swapping Ptet on positive controls.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 78. 9/5/14 (Friday)</span></p><ul class="c5 lst-kix_faphe7ovfpq5-0"><li class="c0"><span>Plan to do: Take cloning through transformation</span></li><li class="c0"><span>Got done: Gel, Digestion, Purification, Ligation, Transformation</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 79. 9/6/14 (Saturday)</span></p><ul class="c5 lst-kix_faphe7ovfpq5-0"><li class="c0"><span>Plan to do: Start liquid cultues</span></li><li class="c0"><span>Got done: Wrapped plates in the morning. Start liquid cultures at night</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 80. 9/7/14 (Sunday)</span></p><ul class="c5 lst-kix_faphe7ovfpq5-0"><li class="c0"><span>Plan to do: Miniprep, Sequencing PCR, Prep for sequencing.</span></li><li class="c0"><span>Got done: Miniprep, Sequencing PCR, Prepare for sequencing and send off tomorrow morning.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 81. 9/10/14 (Wednesday)</span></p><ul class="c5 lst-kix_faphe7ovfpq5-0"><li class="c0"><span>Plan to do: Check on sequences</span></li><li class="c0"><span>Got done: Verified sequence for Bba_pos_moon. Started liquid cultures for experimentation tomorrow. Start designing primers for biobricking.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3"><span class="c2">Day 82. 9/11/14 (Thursday)</span></p><ul class="c5 lst-kix_faphe7ovfpq5-0"><li class="c0"><span>Plan to do: Check for fluorescence of new pTet. Finish and clarify design of primers for biobricking.</span></li><li class="c0"><span>Got done: Bba_pos_moon fluoresces as expected!!!!! Designed primers to start biobricking. Sent for ordering.</span></li></ul><p class="c3 c7"><span></span></p><p class="c3 c7"><span></span></p><p class="c3 c7"><span></span></p><p class="c3 c7"><span></span></p><p class="c3 c7"><span></span></p><p class="c3 c7"><span></span></p><p class="c3 c7"><span></span></p><p class="c7 c14"><span></span></p></body>
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