E. coli transformation with CaCl2

make 5ml  O/N culture (OD around 4). Inoculate a new 5 ml LB with 50 ml O/N culture (OD about 0.05). Let grow again at 37 °C until the new culture becomes "dense" (OD about 0.6). Take 2 ml of this culture for 1 transformation.
 

  • 2 ml LB culture on ice, 5 min
  • Centrifuge 10 sec
  • Remove supernatant
  • Resuspend in 1 ml 100 mM CaCl2
  • 30 min on ice
  • Centrifuge 10 sec
  • Remove supernatant
  • Resuspend in 100 ml 100 mM CaCl2
  • 1 hr on ice
  • add 1-5 ml DNA
  • 30 min on ice
  • 30 sec 42 °C
  • on ice
  • add 600 ml LB
  • 1 hr at 37 °C

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