User:Tamra L. Fisher/Notebook/Research for Dr. Hartings/2012/05/18
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There was no growth on any of the plates from yesterday's transformation. When I removed the microcentrifuge tubes with cells from the incubator shaker, I did notice the cell solutions had begun to turn a bit brownish (which usually is what happens when I leave a tube of leftover cells out overnight). This may be a sign that the cells "went bad" or something in the mixture changed and is affecting them. I thought the tube of cells I used was a new tube of NovaBlue E.coli (not previously defrosted), but maybe not if this happened. I will try the transformations again with another tubes of these cells. This also means we do not have any conclusions on whether or not the PCR from Wednesday was effective or not, since the mini-prepped plasmids should have transformed with no problem.
Sonication of Expressed Triple Mutant Asc Hb in E.coli
All of the transformations from yesterday need to be redone. This includes the Asc Hb mutations M33S, M33S/M103S, and M8S/M33S/M103S into NovaBlue E.coli, the wild-type plamids for Asc Hb and myoglobin (wt swMb pT7-7) into NovaBlue E.coli, and the PCR product from two days ago (Asc Hb M8S/M33S/M103S/A71M) into NovaBlue E.coli. The procedure will be the same as yesterday: