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Team:BYU Provo/Team OxyR/Week18
From 2011.igem.org
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22 August 2011
Matt performed the ligations with the fragments from August 18th. Transformed and plated them, for a long night in the 37˚C incubator.
23 August 2011
Rob and Matt set up colony PCR for the previous day's ligations/transformations. Used primers from the low-copy plasmid backbone, and the GFP gene present in each insert. Rob re-plated the test colonies.
24 August 2011
Rob ran the previous day's colony PCR on gels. We were very excited to see the extremely positive results. Almost every ligation had given at least a few correct colonies... even the ligations from unresolved bands.
Rob started overnight cultures with the positive hits.
25 August 2011
Matt froze down the positive hits in the -80˚C freezer with some DMSO. Our team kept getting better with database management and storage.
Later in the day, Mackay and Matt attempted a plate reader experiment with the new low-copy version of each construct in the DH5α used in the transformation. Mistakes were made ... as so often happens. Matt forgot to make several dilutions, so after the first 1.5 hour stage, they were left with only 25% of the tubes they needed. So, they diluted the already grown tubes into 3 more, further diluting the already dilute mixtures by 4-fold. What with the various complications and mistakes, the results were less than interesting. We noted the problems in the protocol, and prepared a new version for the next time around.
