Team:WashU/Week10

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Revision as of 00:23, 2 August 2012

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

Back to Weekly Log

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-Today we conducted a colony PCR of the TOPO cloning of ZCD with the fusion protein (without a his-tag).  Colony PCR suggested 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.
+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.
 <img src="https://static.igem.org/mediawiki/igem.org/b/b6/Colonyz.jpg">
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