Team:BCCS-Bristol/Protocols-Transformation with heat shock

Transformation with heat shock

1. Thaw the chemical competent bacteria (50 µl E. coli DH5α) on ice
2. Add the DNA by pipetting it directly in the middle of the bacteria solution und mix by stirring gently with the pipette tip
3. Incubate 20 min on ice
4. Heat shock: incubate 60 s at 42°C in a water bath
5. Incubate 2 min on ice
6. Add 250 µl SOC broth (stored in the freezer; thaw before and check for no contamination!)
7. Incubate for 1 h at 37°C 225 rpm
8. Plate 50 µl and 100 µl on two LB plates with antibiotics
9. Incubate the plates at 37°C overnight. Store the remaining bacteria in the fridge.