NGM plates (per Liter)
| Reagent | Final concentration | Amount |
|---|---|---|
| CaCl2 | 1.0 mM | 0.111 g |
| NaCl | 51.3 mM | 3.0 g |
| NH4Cl | 18.7 mM | 1.0 g |
| KH2PO4 | 19.9 mM | 2.7 g |
| K2HPO4 | 5.1 mM | 0.89 g |
| MgSO4 | 1.0 mM | 0.12 g |
| Cholesterol | 130 μM | 5 mg |
| Bacto Peptone | 2.5 g/L | 2.5 g |
| Granulated Agar | 15 g/L | 15 g |
| dH2O | N/A | 1 L |
Prepare stock solutions of 1 M CaCl2, 1M MgSO4, and 1M KPO4 buffer pH6.0 108 3g KH2PO4 and 35.6 g K2HPO4 in 1 L diH2O solution). Autoclave. Prepare 5 mg/mL cholesterol in ethanol. Filter sterilize. Store all of the above solutions at room temperature. In a flask, weight 15 g granulated agar, 2.5 g of Bacto peptone, and 3.0 g of NaCl, add 1 L of diH2O, mix well, autoclave. When the autoclaved solution is cooled to 55°C, add 1 mL 1 M CaCl2, 1M MgSO4, 25 mL 1 M KPO4 buffer PH6.0, and 1 mL 5 mg/mL cholesterol in ethanol to the flask. Mix well. Pour the liquid to 6 cm petri dishes. This media can be stored in a closed plastic box at 22°C–24°C for one month.