Trinh:Chemically Competent Cells

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BE SURE TO PERFORM THIS IN A STERILE ENVIRONMENT. READ AND PREPARE BEFORE PERFORMING.

  • Inoculate 10 ml of SOB medium with seed stock and grow at 37°C to an OD600nm of 0.5
  • Place inoculum on ice for 30 minutes
    • Shake frequently to maintain homogeneous solution
  • Centrifuge at 4000rpm at 4°C for 10 minutes.
  • Discard supernatant by pouring out slowly and pipeting remaining supernatant.
  • Gently resuspend in 10 ml of ice cold CCMB80 buffer.
  • Incubate on ice 20 minutes
  • Centrifuge again at 4°C and discard supernatant as described above.
  • Resuspend in 10 ml of ice cold CCMB80 buffer to an OD of ~3.0
  • Aliquot into chilled screw top or 1.5 mL graduated tubes
  • Store at -80°C.



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