Paris/July 13

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Preparation of Competent Cells

  • Dilution 1/200e (1mL of the O/N 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 unexpected (problem with DNA lyophilized)
  • Transformation of the Biobricks with the "classical" protocol (--> heat shock at 42°C durin 40")
  • Incubation O/N at 37°C of the transformed bacteria


Remark :

  • IGEM 2008 Biobricks : use of the maximum recoverable volume minipreps
  • IGEM 2007 Biobricks : use of 3µl of minipreps
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