Medium data
Search for medium no. = [1135].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
1135 METHANOBACTERIUM MEDIUM FOR STRAIN CAN
| NaCl | 0.5 | g |
| NH4Cl | 0.5 | g |
| K2HPO4 | 0.1 | g |
| KH2PO4 | 0.1 | g |
| CaCl2·2H2O | 0.1 | g |
| MgSO4·7H2O | 0.1 | g |
| FeCl2 solution (see Medium No. 187) | 1.0 | ml |
| Trace element solution (see Medium No. 187) | 1.0 | ml |
| Sodium formate | 2.0 | g |
| Sodium acetate | 1.0 | g |
| FeSO4·7H2O | 0.002 | g |
| L-Cysteine·HCl·H2O | 0.1 | g |
| Resazurin | 0.5 | mg |
| Distilled water | 1.0 | L |
Mix components thoroughly, bring to a boil for several seconds, and cool down under a stream of N2-CO2 (80:20, v/v). Distribute the medium into culture vessels (e.g., 5 ml in the Hungate tube) under the same gas mixture, seal with butyl rubber stoppers and autoclave. Add per liter the following sterile solutions from anaerobic stocks (autoclaved or filter-sterilized):
| 30% Tris base solution | 10.0 | ml |
| 8% Na2CO3 solution | 20.0 | ml |
| Trace vitamins (see Medium No. 197) | 10.0 | ml |
Prior to inoculation, replace the gas phase with a gas mixture of H2-CO2 (80:20, v/v), and add per liter the following solution (autoclaved and stored under a N2 atmosphere):
| 5% Na2S·9H2O solution | 8.0 | ml |
Pressurize the inoculated culture vessels to 200 kPa H2-CO2 (80:20, v/v).
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