Medium data
Search for medium no. = [945].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
945 MB MEDIUM FOR METHANOBACTERIUM
| KH2PO4 | 0.15 | g |
| K2HPO4 | 0.3 | g |
| NaCl | 0.6 | g |
| (NH4)2SO4 | 0.3 | g |
| MgSO4·7H2O | 0.12 | g |
| CaCl2·2H2O | 0.08 | g |
| Trace minerals solution (see below) | 10.0 | ml |
| Resazurin | 1.0 | mg |
| Distilled water | 932.0 | ml |
Mix components thoroughly, and autoclave under a N2-CO2 (80:20, v/v) gas atmosphere. After cooling, aseptically and anaerobically add the following solutions (filter-sterilized):
| 8% NaHCO3 solution | 48.0 | ml |
| Trace vitamins (see Medium No. 197) | 10.0 | ml |
Distribute the medium into culture vessels (leave >80% gas phase by volume) under a H2-CO2 (80:20, v/v) gas mixture, seal with butyl rubber stoppers. Finally, add per liter the following solutions from anaerobic stocks (autoclaved and stored under a N2 atmosphere):
| 5% L-Cysteine·HCl·H2O solution | 6.0 | ml |
| 5% Na2S·9H2O solution | 6.0 | ml |
Pressurize the inoculated vessels to 200 kPa H2-CO2 (80:20, v/v).
Trace minerals solution:
| Nitrilotriacetic acid | 1.5 | g |
| MgSO4·7H2O | 3.0 | g |
| MnSO4·xH2O | 0.5 | g |
| FeSO4·7H2O | 0.1 | g |
| CoSO4·7H2O | 0.1 | g |
| ZnSO4·7H2O | 0.1 | g |
| CuSO4·5H2O | 0.01 | g |
| H3BO3 | 0.01 | g |
| Na2MoO4·2H2O | 0.01 | g |
| Distilled water | 1.0 | L |
Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals. Adjust final pH to 7.0.
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