Paris/July 13

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Preparation of Competent Cells

  • Dilution 1/200e (1mL of culture on 200mL of LB)
  • Culture at 37°C until DO=0,57 (around 3h)
  • Sharing into 4 Falcon tubes of 50mL and cooling at 4°C during 40' in ice
  • Spin during 8', 2000rpm, 4°C
  • Resuspension of the cell's pellet in 20mL of cold CaCl2 (0,1M) during 30' in ice
  • Spin during 8', 2000rpm, 4°C
  • Resuspension of the cell's pellet in 1mL of cold CaCl2 (0,1M)
  • 1h of delay unexcepted (problem with lyophilized DNA)