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Team:BCCS-Bristol/Protocols-Transformation with heat shock
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Latest revision as of 09:49, 14 September 2008
Transformation with heat shock
- Thaw the chemical competent bacteria (50 µl E. coli DH5α) on ice
- Add the DNA by pipetting it directly in the middle of the bacteria solution und mix by stirring gently with the pipette tip
- Incubate 20 min on ice
- Heat shock: incubate 60 s at 42°C in a water bath
- Incubate 2 min on ice
- Add 250 µl SOC broth (stored in the freezer; thaw before and check for no contamination!)
- Incubate for 1 h at 37°C 225 rpm
- Plate 50 µl and 100 µl on two LB plates with antibiotics
- Incubate the plates at 37°C overnight. Store the remaining bacteria in the fridge.
