Team:SDU-Denmark/Protocols/Competent-cells

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Protocol for making competent cells (E.coli)

  # 600 µl overnight (ON) Top10 E. coli culture is added to 60 ml Luria-Bertani (LB) medium.
  # Grows at 37º C while being shaken until the optic density (OD550) is 0,2.
  # Cool cells on ice.
  # Harvest the cells in screwcap tubes (4 × 10 ml).
  # Pour away the supernatant and keep the pellet on ice.
  # Wash the cells with 10 ml cold 50mM CaCl2.
  # The cells are being distributed in eppendorf tubes of 200 µl.
  # Add 41.7 µl 87% glycerol and mix well.
  # Store at -80 º C.