M9 Minimal Media Preparation

1. Dissolve the following salts in about 500mL of ddH2O:
   • 12.8 grams anhydrous Na2HPO4
   • 3.0 grams KH2PO4
   • 0.5 grams NaCl
   • 1.0 gram NH4Cl
2. Bring the media to approximately 950 ml with ddH2O and pH to 7.4 with NaOH.
3. Autoclave for 20 minutes on slow exhaust. Store at 4°C when finished.
4. The following salts were dissolved in 50mL of water and cold filtered with a sterilized 0.22 micron filter:
• 10.8 grams glucose
• 0.19g of MgCl2
• 0.0152 grams CaCl2.2H2O
• 0.0100 grams Thiamine
• 0.007g of FeCl2.4H2O
5. Add the filter sterilized solution into the autoclaved media along with any other additives (e.g. antibiotics). Store at 4°C.