User:Eric M. Walters/Notebook/Spring 2012/2012/05/03

Cyanobacterial acrylate production

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Colony PCR of putative acsA::accBCDA Shit don't segregate, gotta do internal or else you see all your wt copies. Negative control: gDNA Colony PCR reactions and conditions 20 μL reactions: 10 μL GoTaq 1 μL accBCDA-F 1 μL acsA-dnR Either 1 μL gDNA, or pick of colony 7 μL H2O PCR cycle: 95°C - 2:00 95°C - 0:30 61°C - 0:30 72°C - 5:10 (5 kb at 1 kb/min) Repeat 30x 72°C - 10:00 Results Fuckin' nothing