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Team:Cornell/Notebook/Wetlab
From 2012.igem.org
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June
June 10th-16th
June 13th, Wednesday
- Set up PCR for four fragments of the nah operon as well as the pBMT-1 backbone, using primers designed to mutate cut-sites concurrently with Gibson Assembly. Also set up a PCR for the entire nah operon. Due to length of the fragments, a longer extension time was chosen.
- If Gibson Assembly fails, but we are able to PCR the entire 10kb nah operon, an alternate method of mutation using three sequential site-directed mutageneses will be pursued.
- Work done by: Caleb, Claire, Dylan, Spencer, Steven, and Swati
June 14th, Thursday
- PCR only amplified nah1, nah3, and nah4 - longer fragments (nah2, the full nah operon, and pBMT-1) were not identified on the PCR product gel. Set up another PCR with shorter extension time.
June 15th, Friday
- PCR of four out of five products visible on gel. Set up PCR of pBMT-1, final fragment required for Gibson Assembly of the nah operon.
June 17th-23rd
June 24th-30th
June 28th, Wednesday
- Gel purified arsR construct
July
=August=
