IGEM:UC Berkeley/2006/Knockouts
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Procedure for Knockouts
- 50mL LB+Carb+Kan and 5mL starter culture plus 200mg/mL Arabinose
- Grow to OD=.8 (the higher the better)
- Spin approx. 40mL in 50mL conical, 3 minutes @ 8000
- Resuspend 30mL 10% glycerol
- Spin 3 minutes @ 7500
- Repeat glycerol suspension (step 4)
- Dump supernatant, resuspend in remaining liquid (don't add anything in this step)
- Electroporate 50ul cells and 1ul PCR product
- Rescue one hour @ 37C
- Plate