Performed colony PCRs on the 4 sets of cloning reactions that were performed yesterday using the M13F/R vector primers. Colonies were picked, restreaked on a fresh LB Kan50 plates (made 20110726 by SJW) and PCR’d. Master mix calcs are here. Selected 8 white colonies from each cloning reaction for PCR. Restreaked plate was incubated @ 37C O/N.
Cycling Params:
- 95C - 10m
40cycles of:
95C - 10s
55C - 10s
72C - 3m
Results:
Hyperladder I is used as the ladder in both gels.
Cloning results look great (except Colony #1 in the 5’ Top Band didn’t produce a product). Will select a re-streaked colony from each set and inoculate liquid culture for mini prep and subsequent sequencing.