iGEM 2012 — Team Alberta

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# Team:Alberta/Notebook

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iGem Notebook

May

May.9-11.2012

Rick and Tom began coming in to the lab regularly and started learning basic laboratory protocols, such as making competent cells, restriction digests, cell transformation, gel electrophoresis, ect.

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.

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June

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. The competent cell procedure is described elsewhere in this wiki.

June 6

The competent cells were tested with basic puc19 transformation, and the transformation worked with acceptable efficiency.

June 7-8

Today multiple PCRs were run on puc19 with two of the color genes and a repressor gene.

June 11

Made pUC19 plasmid with various color genes and C1 and transformed it into cells.

June 18-21

We cloned a variety of different promoters (nine total) into our color gene plasmids in order to get a sense of relative promoter strengths. We also sent off to sequencing those construct to check that they are correct.

June 22- July 3

We used PCR to add new RBS colour genes and regulatory promoters to our constructs. We then pieced those constructs into kanamycin resistant plasmid backbone and transform them into Top 10 cells.

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July

July 4-5

Torrin and Sarah join the team. We performed the same work on the TG1 strain of E.Coli as a learning exercise.

July 6

Creation of a plasmid with a cut site near the origin of replication began today. This plasmid will be used in order to modify the promoter on the plasmid's origin of replication.

July 9-10

Today Tom Used the successful origin cut site plasmid with Amp Resistance and replaced that resistance with Chlr.

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