Endy:M9 media/supplemented

Component Concentrations

• 1X M9 salts
• 1mM thiamine hydrochloride
• 0.4% glycerol
• 0.2% casamino acids
• 2mM MgSO₄
• 0.1mM CaCl₂

Protocol

For 1L of 1X media

• 500 mL 2X M9 minimal salts ∗
• 30 mL 10 mg/mL thiamine

1. Dissolve 10 mg per mL of H₂O
2. Use a 0.22 μm filter to filter-sterilize
3. Light-sensitive: store covered

• 10 mL 40% glycerol ∗
• 20 mL 10% Casamino acids ∗
• 20 mL 0.1M MgSO₄ ∗
• 200 μL 0.5M CaCl₂ ∗
• 419.8 mL 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 in the media room