User:Karmella Haynes/Notebook/ASU courses/2012/02/14

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Date

  • (BME 494) Starter cultures for Thursday class; 1 section, 9 groups



Plan
> Three experiments comparing Switches A and B

  1. IPTG steady state dose (0 - 1.0 mM) vs. GFP output, liquid
  2. IPTG steady state dose (0 - 1.0 mM) vs. GFP output, agar spots imaging
  3. Time (0 - 18 hrs) with 1.0 mM IPTG vs. GFP output

> Class starts at 3 pm on Thursday (2/16)

  • Two days ahead, starter cultures
  • Day before, experimental cultures (morning)
    • Start Steady State expt. at 6 pm (21 hrs before class)
    • Make spot plates at 9 pm (using Steady State liq samples)
    • Start Time Course expt. (for "final" time point only) at 9 pm (18 hrs.)


Starter cultures
> Overnight starter cultures from agar, 5 mL each, untreated

  1. O2-GFP - amp
  2. O1-GFP - amp
  3. LacI - kan
  4. O2-GFP/ LacI (Switch A) – amp/kan
  5. O1-GFP/ LacI (Switch B) – amp/kan

> Prep enough liquid medium: Will need at least 240 mL Amp/Kan, 10 mL Amp, 10 mL Kan


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