"
Team:Alberta/Notebook
From 2012.igem.org
|
|
iGen Diary
May.9-11.2012
|Rick and Tom recruited and learning basic laboratory protocols, such as making competent cells, restriction digests, cell transformation, gel electrophoresis.
|-
|May. 24
|-
|Rick, Tom, Spencer, and Easwar learned how to use PCR today, with varying degrees of success.
|-
|May 31
|-
|Today Tom finally managed to get a PCR to work, though it took him about twelve attempts.
|-
|June 4
|-
|Made working stock of puc19 and then transformed Top10 with puc19. Overnight cultures were also set up in preparation for the making of competent cells
|-
|June 5
|-
|Competent cells were made today using a standard procedure which took the entire day.
|-
|June 6
|-
|The competent cells were tested with basic puc19 transformation, and the transformation worked.
|-
|June 7-8
|-
|Today multiple PCRs were run on puc19 with two of the color genes and a C1 gene.
|-
|June 11
|-
|Made pUC19 plasmid with the new color genes and C1 and transformed it into cells.
|-
|June 18-21
|-
|We cloned a variety of different (9) promoters into our color gene plasmids, in order to get a sense of relative promoter strengths. we also seqencing those construct to check they are correct
|-
|June 22- july 3
|-
|We PCR new RBS colour genes and regulatory promoters. we then made those constructs into kanamycin resistant plasmid backbone and transform them into Top 10 cells.
|-
|July 4-5
|-
|Torrin and Sarah Join. we perform the same work again on TG1.
|-
|July 6
|-
|Tom checked overnights of origin cut site strains and chose colonies to be sequenced.
|-
|July 9-10
|-
|Today Tom Used the successful origin cut site plasmid with Amp Resistance and replaced that resistance with Chlr.
Torrin and Sarah are creating and testing chemical gradient plates, and learning additional laboratory procedure.
Rick and Spencer is making competent cells of TG1 so that we can test our current parts in an E.Coli strain that grows faster than our current Top10.
|-
|Rick and Easwar are drop in LacI and Tet R repressor into chloranphenicol construct.
|-
|July 11
|-
|We continued our experiments with making chemical gradients on agar plates.
|-
|
|-
|
