Team:Cambridge/Protocols/CCMB80

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CCMB80 buffer Recipe

This buffer is used when making chemically competent cells

  • 10 mM KOAc pH 7.0 (10 ml of a 1M stock/L or 0.98g/L)
  • 80 mM CaCl2.2H2O (11.8 g/L)
  • 20 mM MnCl2.4H2O (4.0 g/L)
  • 10 mM MgCl2.6H2O (2.0 g/L or 5ml of 2M stock/L)
  • 10% glycerol (100 ml/L)
  • Fill up with DI water AFTER regulating pH
  • adjust pH DOWN to 6.4 with 0.1N HCl if necessary
    • adjusting pH up will precipitate manganese dioxide from Mn containing solutions.
  • sterile filter and store at 4°C
  • slight dark precipitate appears not to affect its function