Overview
Week 1
Week 2
Week 3
Week 4
Week 5
Week 6
Week 7
Week 8
Week 9
Week 10
Week 11
Week 12
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Week Eight |
Monday, 8/4/14
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Export Systems
Attempt to de-FLAG pTG004 to create lam system in its "native state" for use in LCMS:
- Ran a colony PCR on 4 colonies picked from a CARB plate that had been plated with a 2nd Round-the-horn reaction.
- A gel was run on the products, and a liquid culture was set up for the one colony suggesting it contains a properly formed pTG004-noFLAG plasmid.
Attempt to create a negative control for Western blotting:
- Reattempted transformation of Anton's B2 pTet_GFP GG-assemblies into JM109. Plated on Cm plates & incubated overnight.
agrBCDA Reception System and Combinatorial Promoters
- Transformed JM109 cells with the pAA009 & pAA010 Gibson assembly products from Friday.
- Transformed DH5α-Z1 cells with the pAA002 Gibson assembly products from Friday.
Beginning of a new, TXTL characterization project:
- We began characterization of a constitutive promoter family designed by Berkeley iGEM 2006 (biobrick parts BBa_J23100-J23118). We transformed JM109 cells with the re-suspended plasmids.
- Some parts were not included with the kit mailed to us, so we might request those at a later date.
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Tuesday, 8/5/14
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Export Systems
lamBCDA & fsrABC Reception Systems
- Reran colony PCR using the new primers that we ordered.
agrBCDA Reception System and Combinatorial Promoters
agrBCDA system:
- Only 1 colony was resulted from yesterday's transformation of pAA009. Set up a liquid culture of this colony.
- Ran gradient PCR on pAA009 backbone using 6 temperatures from 50-70°C.
- DpnI digested pAA009 backbone for 2 hours and stored at 4°C overnight.
Combinatorial promoters project:
- Picked 6 colonies from the plate transformed yesterday with pAA002 and set up liquid cultures.
TXTL characterization project:
- Miniprepped transformed liquid cultures grown up last night
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Wednesday, 8/6/14
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Export Systems
- Set up overnight liquid culture of pMB006-transformed colony that looks promising based on yesterday's colPCR
lamBCDA & fsrABC Reception Systems
agrBCDA Reception System and Combinatorial Promoters
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Thursday, 8/7/14
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Export Systems
lamBCDA & fsrABC Reception Systems
- Miniprepped overnight liquid culture of pMB006. Shipped to Eurofin for sequencing.
agrBCDA Reception System and Combinatorial Promoters
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Friday, 8/8/14
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Export Systems
lamBCDA & fsrABC Reception Systems
- Analyzed sequencing results. They were messy and revealed nothing about the sequence.
agrBCDA Reception System and Combinatorial Promoters
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