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Team:Paris Liliane Bettencourt/Notebook/2011/08/15/
From 2011.igem.org
Contents |
Kevin
Spread transformated cells
Because of uniform growing, it wasn't possbile to select one clone. So we spread bacteria on new plates, overnight.
Cyrille
pHM3 miniprep
The clones picked yesterday where grown overnight. The result in term of color was not homogenious at all, and in particular one tube that was incredibly violet. We wanted to understand where this diversity is comming from.
In one of the pale red tube, we add some IPTG, which turn out to make it a bit more red, but not much more.
The second hypothesis is that the very violet plasmid is more efficient than the others. To do that, I prepared 4 replicates ( a, b, c,d ). A few hours later, they turn out to be the same color than the others.
We wondered if the plasmid was not able to do intra recombination , keeping the resistance and loosing the rest. To test that, we tried a few replicates of the very red colony, and make it grow on tetracyclin antibiotic. We didn't obsereved a significant growth after a few hours.
Finally, at the end end of the day, we made two glycerol from two replicates of the very red tube (S69) and centrifugate and freeze the rest for miniprep tomorow
Competent cell preparation
Competent MH1 cells where prepared using the same protocol from than day .
