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User contributions
From 2014.igem.org
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- 20:37, 17 October 2014 (diff | hist) N Harvard BioDesign/28 September 2014 (Created page with "Plated more CsgA--This is growing up on the bench and will be ready for congo red/ incubation on Tuesday (used pHBD48 as neg ctrl because our large yesca plates are only carb--i ...") (top)
- 20:37, 17 October 2014 (diff | hist) N Harvard BioDesign/13 September 2014 (Created page with "Inoculated 50ml cultures of 46 & 48 Inoculated 5ml cultures of PHL -, 53 (Csga), 54 (Csga-spy)") (top)
- 20:36, 17 October 2014 (diff | hist) N Harvard BioDesign/11 September 2014 (Created page with "Purified pHBD48 and pHBD46 Inoculated more pHBD46 and pHBD48") (top)
- 20:36, 17 October 2014 (diff | hist) N Harvard BioDesign/10 September 2014 (Created page with "Made YESCA-IPTG-CRB plates") (top)
- 20:35, 17 October 2014 (diff | hist) Team:Harvard BioDesign/Notebook
- 20:35, 17 October 2014 (diff | hist) N Harvard BioDesign/7 August 2014 (Created page with "Ran SDS Page gel on incubation products of CsgA Spy Tag and Chromoprotein-Spy Catcher Gibsoned parts of pHBD 68, pHBD 69, pHBD 70, pHBD 72, pHBD 73 and pHBD 74 Transformed Gibson...") (top)
- 20:34, 17 October 2014 (diff | hist) N Harvard BioDesign/4 August 2014 (Created page with "Congo red spin down on induced LSR10 cells with SpyTag, SynZIP #1 and SynZIP #5 Incubate cells expressing SpyTag on their curli monomers with Chromoprotein-SpyTag lysate Resul...") (top)
- 20:33, 17 October 2014 (diff | hist) N Harvard BioDesign/1 August 2014 (Created page with "Picked colonies containing pHBD 54, 55, 56, and 57 (CsgA tagged with SpyTag, Synzip 1, Synzip 4, and SynZip 5 respectively) and inoculated them in LSR10s and PHLs. After cells r...") (top)
- 20:32, 17 October 2014 (diff | hist) N Harvard BioDesign/31 July 2014 (Created page with "Sequencing results came back viable for…. SynZIP1_C SynZIP4_A,B,C SynZIP5 (there is a deletion in flag tag, but the insert is there) SpyTag A Received HBD 108-111 (Primers to ...") (top)
- 20:32, 17 October 2014 (diff | hist) N Harvard BioDesign/30 July 2014 (Created page with "Ran gel to verify PCR products that we Gibsoned yesterday Backbone (for SynZIPs)- CsgA_48Linker_Flag Backbone (for SpyTag)- CsgA_48Linker Grow up colonies picked from plates wit...") (top)
- 20:32, 17 October 2014 (diff | hist) N Harvard BioDesign/29 July 2014 (Created page with "Colonies from Gibson of pBbE1a backbone- CsgA- 48 Linkiner- SynZIP1 did not grow Error amplifying the backbone HBD 5 and 7 were used as primers (point inward towards CsgA) Primer...") (top)
- 20:31, 17 October 2014 (diff | hist) N Harvard BioDesign/28 July 2014 (Created page with "Received Synzips 1 and 5 Gibsoned Synzip #1 and Synzip #2 into the CsgA-48 linker backbone Transformed Synzip #1 into Mach I cells * Synzip #5 ← → Synzip #6 * Synzip #1 ...") (top)
- 20:31, 17 October 2014 (diff | hist) N Harvard BioDesign/25 July 2014 (Created page with "Sequencing results The stop codon was missing from the ordered oligonucleotide blocks that were used for sequencing Miniprepped 11 Mach I- SpyTag- CsgA colonies growing up from ...") (top)
- 20:31, 17 October 2014 (diff | hist) N Harvard BioDesign/24 July 2014 (Created page with "Picked colonies from July 23rd transformations to grow up Place in 3 milliliters of LB + carb Picked 11 colonies to grow Add 7 milliliters of LB+ carb to the already growing tr...") (top)
- 20:30, 17 October 2014 (diff | hist) Harvard BioDesign/23 July 2014 (top)
- 20:29, 17 October 2014 (diff | hist) N Harvard BioDesign/23 July 2014 (Created page with "Sequencing results were not good The plasmid did not have the insert")
- 20:28, 17 October 2014 (diff | hist) N Harvard BioDesign/22 July 2014 (Created page with "Waited the whole day for the Mach I cells to grow up, but it seemed unsuccessful") (top)
- 20:28, 17 October 2014 (diff | hist) N Harvard BioDesign/21 July 2014 (Created page with "Grew up colonies from transformation plates: Mach I cells with SpyTag- CsgA Only one of the plates really had colonies. Not quite sure why the growth was so small? Plated 100 mi...") (top)
- 20:26, 17 October 2014 (diff | hist) Harvard BioDesign/18 July 2014 (top)
- 20:26, 17 October 2014 (diff | hist) N Harvard BioDesign/18 July 2014 (Created page with "Picked colonies from pHBD45, pHBD1 and pHBD2 transformations from previous day. Mini-prepped pHBD45 Ran PCR fragments for the gibson of pHBD40 from previous day on a gel, gel e...")
- 20:22, 17 October 2014 (diff | hist) N Harvard BioDesign/17 July 2014 (Created page with "→ No LB/25mL tubes...need to pick colonies tomorrow PCR’ed gibson fragments for pHBD40 with new primers (HBD71-74)") (top)
- 20:22, 17 October 2014 (diff | hist) N Harvard BioDesign/16 July 2014 (Created page with "Received pDEST14_His6_Spycatcher plasmid (pHBD45) from Peter→ transformed into MACH1s and plated to make our own stocks Transformed pHBD1 and pHBD2 into LSR10 & plated for cry...") (top)
- 20:22, 17 October 2014 (diff | hist) N Harvard BioDesign/10 July 2014 (Created page with "Transformed successful Gibsons: Chi C and CBD1_1 into LSR10 cells Grew up Turbo and LSR10 cells for a biocompatibility assay PCRed F12 and F48 backbones to grow stocks for future...") (top)
- 20:21, 17 October 2014 (diff | hist) N Harvard BioDesign/9 July 2014 (Created page with "PCRed plasmid backbones with the F12 and F48 linkers In order to perform more Gibson reactions Ran a gel on the PCRed backbone Assessed results of biocompatibility assay 1 ---...") (top)
- 20:21, 17 October 2014 (diff | hist) Harvard BioDesign/8 July 2014 (top)
- 20:20, 17 October 2014 (diff | hist) N Harvard BioDesign/8 July 2014 (Created page with "Gel extracted parts for HybB + SB3K3 for Gibson. Ran Gibson. Transformed. Picked colonies from PCR of pHBD39-41 to clone out prefix. Only pHBD41 had many colonies. Picked 2 col...")
- 20:19, 17 October 2014 (diff | hist) N Harvard BioDesign/7 July 2014 (Created page with "Ran PCR to remove prefix from pHBD39-41. Gel extraction only gave good concentration for pHBD41. Transformed all into MACH1 cells + plated @ 10uL and 60uL. PCR’ed HybB promot...") (top)
- 20:18, 17 October 2014 (diff | hist) N Harvard BioDesign/2 July 2014 (Created page with "We began minipreps of pHBD 1, 2, 43, and 44. We then ran our PCRs of fragments of pHBD 26 and 28 from yesterday on a gel: pHBD 18 and 35 were successfully PCRed, so those were ...") (top)
- 20:18, 17 October 2014 (diff | hist) N Harvard BioDesign/1 July 2014 (Created page with "Our LacO hypothesis was rejected as none of the IPTG induced cultures showed any coloring. However, we noticed that one of our plates with pHBD44 had blue colonies! Even though ...") (top)
- 20:17, 17 October 2014 (diff | hist) Harvard BioDesign/30 June 2014 (top)
- 20:16, 17 October 2014 (diff | hist) N Harvard BioDesign/30 June 2014 (Created page with "Promoter Cloning (HSP & ASP into SB3K3 backbone): We looked at our cultures that we placed in the 25 C and 30 C shakers over the weekend, but there was no color. We weren’t su...")
- 20:14, 17 October 2014 (diff | hist) N Harvard BioDesign/27 June 2014 (Created page with "We began minipreps on the following: Our retransformations of pHBD 5 and 35 Our gibsons of pHBD 39, 40, 41, 42, and 44. We finished minipreps of all of them. We also innoculated...") (top)
- 20:13, 17 October 2014 (diff | hist) N Harvard BioDesign/26 June 2014 (Created page with "We miniprepped pHBD 21 and 24. We retransformed pHBD 5 and 35. pHBD 37 arc’ed so we decided to pick from the original biobrick plate tomorrow morning when we inoculate culture...") (top)
- 20:13, 17 October 2014 (diff | hist) N Harvard BioDesign/25 June 2014 (Created page with "Finished the minipreps of BBa_K274200, BBa_I765001, pHBD4, BBa_K1073023, BBa_K1033931, and BBa_K1033929. Plated Gibson transformations of pHBD24 and pHBD21 had few (1-2) colonie...") (top)
- 20:12, 17 October 2014 (diff | hist) N Harvard BioDesign/24 June 2014 (Created page with "We picked up more stabs from iGEM HQ. We got three new chromoproteins: BBa_K1073023 (eforRed [pink]), BBa_K1033931 (amilGFP [yellow]), and BBa_K1033929 (aeBlue [indigo]). While w...") (top)
- 20:11, 17 October 2014 (diff | hist) N Harvard BioDesign/23 June 2014 (Created page with "Made electrocompotent MACH1 and PHL cells We ran the pbBE1a backbone from yesterday on a gel and this time we got bands We gel purified the backbone and hybB insert PCR produc...") (top)
- 20:10, 17 October 2014 (diff | hist) N Harvard BioDesign/20 June 2014 (Created page with "We made a 2% agarose gel and ran our PCR products on the gel. Lane 1 is a 1 kb ladder and 8 is a 100 bp ladder. 2 and 3 are both the pbBE1a product, while lanes 4-7 are all the h...") (top)
- 20:10, 17 October 2014 (diff | hist) N Harvard BioDesign/19 June 2014 (Created page with "We ran a PCR on the pbBE1a backbone as well as a colony PCR on a Mach1 in order to isolate the hybB promoter. We also miniprepped the chromoproteins and pSB3K3 (pHBD 32-35) Discu...") (top)
- 20:09, 17 October 2014 (diff | hist) N Harvard BioDesign/18 June 2014 (Created page with "Miniprepped pHBD 13, 15, 16, 18 (13 and 18 biological replicates since concentrations from previous day’s miniprep were very low) pHBD20 didn’t grow Transformed chromoprotei...") (top)
- 20:09, 17 October 2014 (diff | hist) N Harvard BioDesign/17 June 2014 (Created page with "Some plates didn’t have individual colonies; just smears. This is likely due to the fact that we added too little antibiotic to our plates. Instead of adding 250 µl of CM, we ...") (top)
- 20:08, 17 October 2014 (diff | hist) N Harvard BioDesign/16 June 2014 (Created page with "We got our sequencing results and aligned them to our plasmid in geneious. 3 out of 4 of the sequencing reactions worked. We plan to sequence the entirety of pHBD1ta or pHBD1tb (...") (top)
- 20:06, 17 October 2014 (diff | hist) N Harvard BioDesign/13 June 2014 (Created page with "Plates from the Gibson assembly (pBbE1a-tet-CsgA) transformations grew many colonies! We picked 2 Turbo and 2 MACH1 colonies and inoculated in 5mL LB+Carb to shake up for mini-p...") (top)
- 20:05, 17 October 2014 (diff | hist) N Harvard BioDesign/12 June 2014 (Created page with "PCR Primers from John→ HBD 1-4 HBD 1 & 2 = fwd and rev for pBbE1a-CsgA backbone HBD 3 & 4 = fwd and rev for Tet promoter Backbone template from Bom → pBbE1a-CsgA @ 65ng/...") (top)
- 20:05, 17 October 2014 (diff | hist) N Harvard BioDesign/11 June 2014 (Created page with "Preparing electrocompetent cells Picked two 5mL LSR10 inoculated cultures and added them each to a 500mL Low Na LB broth flask. Flasks were left shaking at 37C and periodically ...") (top)
- 20:03, 17 October 2014 (diff | hist) N Harvard BioDesign/10 June 2014 (Created page with "Picking colonies Picked colonies from 200uL plated PUC19 transformed cells, 10uL plated pBbe1a (- ctrl) transformed cells, and 10uL pBbe1a CsgA Arc’ed cells. There were many c...") (top)
- 20:02, 17 October 2014 (diff | hist) Team:Harvard BioDesign/Notebook
- 19:49, 17 October 2014 (diff | hist) N Harvard BioDesign/9 June 2014 (Created page with "Making Plates! YESCA + 100ug/mL Carb + 25ug/mL CR + 3ug/mL CBB + 0.5mM IPTG YESCA + 25ug/mL Cm + 25ug/mL CR + 3ug/mL CBB + 0.5mM IPTG Pure LB LB+ 100ug/mL Carb LB + 15ug/mL Cm...") (top)
- 19:48, 17 October 2014 (diff | hist) Team:Harvard BioDesign/Notebook
- 19:47, 17 October 2014 (diff | hist) Team:Harvard BioDesign/Notebook
- 19:47, 17 October 2014 (diff | hist) Team:Harvard BioDesign
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