Medium data
Search for medium no. = [231].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
231 METHANOBACTERIUM THERMOAUTOTROPHICUM MEDIUM
| KH2PO4 | 0.3 | g |
| (NH4)2SO4 | 1.5 | g |
| NaCl | 0.6 | g |
| MgSO4·7H2O | 0.12 | g |
| CaCl2·2H2O | 0.08 | g |
| FeSO4·7H2O | 4.0 | mg |
| K2HPO4 | 0.15 | g |
| Na2CO3 | 4.0 | g |
| Trace vitamins (see Medium No. 197) | 10.0 | ml |
| Trace element solution (see below) | 10.0 | ml |
| L-Cysteine·HCl·H2O | 0.5 | g |
| Na2S·9H2O | 0.5 | g |
| Resazurin | 1.0 | mg |
| Distilled water | 980.0 | ml |
Mix ingredients, except cysteine·HCl and Na2S·9H2O, and adjust pH to 7.2. Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20). Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-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. Pressurize the inoculated bottles to 100 kPa H2-CO2 (80:20).
Trace element solution:
| EDTA·2Na | 0.64 | g |
| MgSO4·7H2O | 6.2 | g |
| MnSO4·xH2O | 0.55 | g |
| NaCl | 1.0 | g |
| FeSO4·7H2O | 0.1 | g |
| CoCl2·6H2O | 0.17 | g |
| CaCl2·2H2O | 0.13 | g |
| ZnSO4·7H2O | 0.18 | g |
| CuSO4·5H2O | 0.05 | g |
| KAl(SO4)2·12H2O | 0.018 | g |
| H3BO3 | 0.01 | g |
| Na2MoO4·2H2O | 0.011 | g |
| NiCl2·6H2O | 0.025 | g |
Dissolve EDTA·2Na first in 500 ml of distilled water. Then add the remaining salts and adjust volume to 1000 ml.
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