Endy:M9 media/supplemented

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

• 500 ml 2xM9 salts ∗
• 30 mL 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 ∗
• 419.8 ml 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