Endy:M9 media/supplemented

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This is our protocol for making supplemented M9 medium.

Component Concentrations

  • M9 minimal medium containing glycerol as the carbon source supplemented with -
  • 1mM thiamine hydrochloride
  • 0.2% casamino acids

Protocol

For 1L of media

  • 500ml 2xM9 salts
  • 30ml 10 mg/ml thiamine hydrochloride
    1. Dissolve 10 mg per ml of H2O
    2. Filter-sterilize using a 0.22μm filter
    3. Light-sensitive: store covered
  • 10ml 40% glycerol ∗
  • 20ml 10% casamino acids ∗
  • 20ml 0.1M MgSO4
  • 200μl 0.5M CaCl2
  • 419.8ml sterile deionized H2O ∗

Combine above solutions using sterile technique. (May notice some precipitation during preparation but precipitate should go back into solution once volume is brought up to 1l with sterile H2O.)
Add antibiotic as appropriate and store at 4°C

∗ Can be obtained from the media room

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