Medium data
Search for medium no. = [272].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
272 METHANOCALCULUS MEDIUM
| KH2PO4 | 0.4 | g |
| NH4Cl | 0.9 | g |
| MgCl2·6H2O | 0.36 | g |
| NaCl | 10.0 | g |
| Sodium acetate | 0.5 | g |
| Yeast extract (BD-Difco) | 2.0 | g |
| Trypticase peptone (BD-BBL) | 2.0 | g |
| Trace vitamins(see Medium No. 197) | 10.0 | ml |
| Trace element solution (see Medium No. 243) | 10.0 | ml |
| NaHCO3 | 5.0 | g |
| L-Cysteine·HCl·H2O | 0.5 | g |
| Na2S·9H2O | 0.5 | g |
| Resazurin | 1.0 | mg |
| Distilled water | 1.0 | L |
Mix ingredients except NaHCO3 and adjust pH 7.0. Bring to a boil for 5-10 seconds, then, cool down under a stream of H2-CO2 (80:20, v/v) and add NaHCO3. Dispense the medium into suitable culture vessels (e.g. 20 ml in 120 ml serum bottles) under a stream of H2-CO2 (80:20, v/v) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Prior to inoculation, aseptically and anaerobically add per liter the following solutions (autoclaved and stored under a N2 atmosphere):
| 5% Na2S·9H2O solution | 10.0 | ml |
| 5% L-Cysteine·HCl·H2O solution | 10.0 | ml |
Adjust pH to 7.5, if necessary. Pressurize the inoculated bottles to 200kPa H2-CO2 (80:20, v/v).
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