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

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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