Team:Newcastle/E. coli Competence

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Competent cells preparation

  1. Inoculate 300ml with 1/20 volume of an overnight culture of the desired strain.
  2. Grow the cell at 37°C (with shaking)
  3. Chill cells on ice and harvest by centrifugation at 4°C for 10 minutes
  4. Resuspend the pellet in 100ml of ice cold TFBII
  5. Aliquot 200µl volumes of the cell suspension into cooled sterile microfuge tubes and flash freeze in dry-ice
  6. Store at -80°C
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