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Team:Cornell/notebook
From 2014.igem.org
Notebook
Week 1 (June 2 - June 8)
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Wetlab Overview
Transformation efficiency and protocols were tested - heat shock did not work well, and we came to the conclusion to stick with electroporation. The first day of wetlab training began on the 7th! We started working on amplification of metallothionein genes - so far nixA and GST-PMT appear to be working.
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Drylab Overview
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Week 2 (June 9 - June 15)
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Wetlab Overview
Obtained transformants of pA14F and pC14T heat shocks and both E/B and E/D were successfully heat shocked into DH5a. Ran into problems later in the week with heat shocked plates from overgrowth and contamination; switched to electroporating of C+F and E+B.
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Mercury Subteam
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Nickel Subteam
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Reporters Subteam
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Drylab Overview
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Week 3 (June 16 - June 22)
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Wetlab Overview
This week we ligated various parts (F+A, F+C, E+B, and E+D), transformed, ran colony PCRs, and submitted them for sequencing. Most of our attempts were unsuccessful either due to failed ligations or contamination. We also made T for future minipreps.
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Drylab Overview
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Week 4 (June 23 - June 29)
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Wetlab Overview
Cloning and troubleshooting continues. E+D, F+C, E+D, E+B4, and E+D11 sequencing results came in negative. The restriction cocktail method was used on F+C and F+A because mRFP kept religating back into the backbone. Plates were successful, but colony PCRs showed they were negative. Repeat cloning. We are facing problems with contamination of lead transporter and R plate. Submitting F+C3 for sequencing. We are also working on transforming off the AA and AB kit plate.
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Drylab Overview
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Week 5 (June 30 - July 6)
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Wetlab Overview
Transformants gave negative signals; many gels lacked proper bands in correct locations. Growth assay looked good for mercury although concentration needs to be increased for nickel.
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Drylab Overview
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Week 6 (July 7 - July 13)
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Wetlab Overview
We gathered the data for an assay testing the growth of cells in different heavy metals. In addition, we ligated each the lead transporter (R), and nickel (AA) and mercury (AB) inducible promoters with H. Unfortunately, our transformations did not seem to work for reasons like incomplete digests and inefficient stocks.
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Drylab Overview
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Week 7 (July 14 - July 20)
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Wetlab Overview
We are troubleshooting some problems with gel purification and smearing on the gel when we visualize it. Our new stocks of competent cells also appear to be bad. Sequencing of R+H returned a portion of a random promoter region of Enterococcus, so this will have to be remade. We are in the process of creating many different BioBrick parts, such as AB1+H, F, AC+H, AB2+H, AC1S+H, AB2S+H, AC+H, AA + H, which are all at various stages of the cloning process.
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Drylab Overview
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Week 8 (July 21 - July 27)
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Wetlab Overview
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Drylab Overview
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Week 9 (July 28 - August 3)
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Wetlab Overview
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Drylab Overview
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Week 10 (August 4 - August 10)
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Wetlab Overview
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Drylab Overview
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Week 11 (August 11 - 17)
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Wetlab Overview
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Drylab Overview
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Week 12 (August 18 - August 24)
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Wetlab Overview
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Drylab Overview
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Week 13 (August 25 - August 31)
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Wetlab Overview
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Drylab Overview
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Week 14 (September 1 - September 7)
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Wetlab Overview
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Drylab Overview
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Week 15 (September 8 - September 14)
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Wetlab Overview
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Drylab Overview
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Week 16 (September 15 - September 21)
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Wetlab Overview
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Drylab Overview
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Week 17 (September 22 - September 28)
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Wetlab Overview
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Drylab Overview
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Week 18 (September 29 - October 5)
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Wetlab Overview
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Drylab Overview
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Week 19 (October 6 - October 12)
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Wetlab Overview
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Drylab Overview
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Week 20 (October 13 - October 19)
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Wetlab Overview
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Drylab Overview
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