Monday, July 30
Failed PCR in morning
Digestion of Z construct and tet-resistant plasmid with E and S. Run on gel
PCR using KlenTaq - worked
Transformed GC5 cells with Z-construct and U Plated cells to check tomorrow
Tuesday, July 21
Colony PCR on TOPO Ligation of Z construct to T plasmid PCR of Z and U constructs
Wednesday, August 1 Today we conducted a colony PCR of the TOPO cloning of ZCD with the fusion protein (without a his-tag). Colony PCR showed a single band of approximately 1200bp for the sample of colony number two. A successful colony PCR should yeild amplificaiton of ZCD with a 60bp upstream region, for a total of 1170bp. The close agrement between the band size and the expected band suggest that colony number 2 had the correct orientation of the insert (gel below). This colony will be grown up overnight and minipreped in the morning. Thursday, August 2
Friday, August 3
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