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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