Date
- (BME 494) Starter cultures for Thursday class; 1 section, 9 groups
Plan
> Three experiments comparing Switches A and B
- IPTG steady state dose (0 - 1.0 mM) vs. GFP output, liquid
- IPTG steady state dose (0 - 1.0 mM) vs. GFP output, agar spots imaging
- 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
- O2-GFP - amp
- O1-GFP - amp
- LacI - kan
- O2-GFP/ LacI (Switch A) – amp/kan
- 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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