Medium data
Search for medium no. = [1039].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
1039 METHANOBACTERIUM MEDIUM (VII)
| KH2PO4 | 0.5 | g |
| MgSO4·7H2O | 0.4 | g |
| NaCl | 0.4 | g |
| NH4Cl | 0.4 | g |
| CaCl2·2H2O | 0.05 | g |
| FeSO4·7H2O | 2.0 | mg |
| FeCl2 solution (see Medium No. 187) | 1.0 | ml |
| Trace element solution (see Medium No. 187) | 1.0 | ml |
| Brain heart infusion (BD-Difco) | 6.0 | g |
| Proteose peptone (BD-Difco) | 6.0 | g |
| Yeast extract | 2.0 | g |
| Sodium acetate | 1.0 | g |
| Sodium formate | 2.0 | g |
| NaHCO3 | 4.0 | g |
| Resazurin | 1.0 | mg |
| Distilled water | 1.0 | L |
Mix components, except NaHCO3 and vitamin solution, and bring to a boil for 5-10 sec. Cool down under a stream of H2-CO2 (80:20, v/v) and add NaHCO3 to the medium. Dispense the medium into culture vessels under a stream of H2-CO2 (80:20, v/v), seal with butyl rubber stoppers and autoclave. Prior to inoculation, add per liter the following solutions (autoclaved or *filter-sterilized and stored under a N2 atmosphere).
| Vitamin solution* (see below) | 1.0 | ml |
| 5% Na2S·9H2O solution | 10.0 | ml |
| 5% L-Cysteine·HCl·H2O solution | 10.0 | ml |
After growth has started and the culture is becoming turbid, pressurize to 50-100 kPa H2-CO2 (80:20, v/v).
Vitamin solution:
| Vitamin B12 | 100.0 | mg |
| p-Aminobenzoic acid | 80.0 | mg |
| Biotin | 20.0 | mg |
| Nicotinic acid | 200.0 | mg |
| DL-Calcium pantothenate | 100.0 | mg |
| Pyridoxine·HCl | 300.0 | mg |
| Thiamine·HCl | 200.0 | mg |
| Distilled water | 1.0 | L |
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