Biology Protocols

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Minimal Medium

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Stock Solutions

M9 minimal salts solution (5X concentrate)

  • To 800mL of distilled/deionized water add:
  • 64g Na2HPO4-7H2O
  • 15g KH2PO4
  • 2.5g NaCl
  • 5.0g NH4Cl
  • Make to 1 litre with dH20
  • Sterilze by autoclaving or filter sterilization if autoclave is not available.

1 M MgSO4

  • 24.65 g MgSO47H2O
  • Water to make 100 ml

40% glucose (w/v)

  • 40g Glucose
  • Water to 100ml
  • Add glucose to stirring water in beaker; DO NOT ATTEMPT TO ADD WATER TO GLUCOSE!
  • Stir until all dissolved — may take 45 minutes or more of stirring.

1M CaCl2

  • 147.014g CaCl2·2dH20
  • Make to 1 litre with dH20


To prepare 1L of media, add:

  • 200mL 5X M9 salts solution
  • to ~800mL of distilled water
  • Add 15g of agar media if agar plates are to be poured.

After autoclaving, swirl to mix evenly and "temper" at room temperature (until you can place your hand on the flask for 2 second) then add:

  • 2mL of 1M MgSO4 solution
  • 0.1mL of 1M CaCl2 solution
  • 20mL of 20% glucose

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