User:Eric M. Walters/Notebook/Spring 2012/2012/05/31

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Enriching for barcode KO

  • So the 12 colonies from yesterday all appear to have the barcode replacing at least one acsA. Hell yeah. Now I need to select for more loci being replaced. Doing that by making 12 patches on 5 mM AA plates. Gotta make those plates first (making a 250 mL batch). Rewriting the protocol here because I keep just looking back to old stuff.

5mM AA plates

  • Autoclave 125 mL of 2x Media A (in the 4), and 125 mL of 6% Bacto agar (3.75 g)
  • Cool it a bit in the 50 bath
  • Add: 12.5 uL B12, 2.5 mL .5M AA stock