User:Karmella Haynes/Notebook/ASU courses/2012/02/15

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02/15/12

  • (BME 494) Experimental cultures and plates



Experiment specs
> NEB10B cells
> Incubations at 37 °C
> LB Lennox media buffered with 1 mL/L NaOH


Experimental cultures for Monday sections
> Morning: Inoculate cultures w/ 100 uL starter culture (30 tubes total, 10 mL each)

Experiment 1: Dose Response, liquid
> 15 tubes, start at 9 pm

  Medium IPTG (mM)
O2-GFP (1) Amp           1.0
O2-GFP/ LacI-O2 (2-7) Amp/ Kan 0.0 0.05 0.1 0.25 0.5 1.0
O1-GFP (8) Amp           1.0
O1-GFP/ LacI-O2 (9-14) Amp/ Kan 0.0 0.05 0.1 0.25 0.5 1.0
LacI-O2 (15) Kan           1.0


Experiment 3: Time course
> 15 tubes

  Medium Time to add 1.0 mM IPTG
O2-GFP Amp 9pm          
O2-GFP/ LacI-O2 Amp/ Kan 9pm 6am 8am 10am 12pm none
O1-GFP Amp 9pm          
O1-GFP/ LacI-O2 Amp/ Kan 9pm 6am 8am 10am 12pm none
LacI-O2 Kan 9pm         none


2/16/12 (morning)

Experiment 2: Dose response, agar
> Pipette 5 μL of Experiment 1 samples onto agar
> Amp/ Kan: Experiment 1 samples 2-7 and 9-14 (3 plates)
> Amp: Experiment 1 samples 1 and 8 (3 plates)

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