Medium data
Search for medium no. = [230].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
230 METHANOSARCINA MEDIUM
| KH2PO4 | 0.227 | g |
| K2HPO4 | 0.348 | g |
| NH4Cl | 0.5 | g |
| MgSO4·7H2O | 0.5 | g |
| CaCl2·2H2O | 0.25 | g |
| NaCl | 2.25 | g |
| FeSO4·7H2O | 2.0 | mg |
| Yeast extract (BD-Difco) | 2.0 | g |
| Casitone (BD-Difco) | 2.0 | g |
| Trace vitamins (see Medium No. 197) | 10.0 | ml |
| FeCl2 solution (see Medium No. 187) | 1.0 | ml |
| Trace element solution (see Medium No. 187) | 1.0 | ml |
| NaHCO3 | 0.85 | g |
| Methanol | 10.0 | ml |
| L-Cysteine·HCl·H2O | 0.3 | g |
| Na2S·9H2O | 0.3 | g |
| Resazurin | 1.0 | mg |
| Distilled water | 980.0 | ml |
Mix ingredients, except NaHCO3, methanol, cysteine·HCl and Na2S·9H2O, and bring to a boil for 5-10 sec. Cool down under a stream of N2-CO2 (80:20), and add NaHCO3 and methanol to the medium. Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of N2-CO2 (80:20) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Separately autoclave cysteine·HCl and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the cysteine·HCl and Na2S·9H2O solutions.
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