Medium data
Search for medium no. = [1331].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
1331 HYDROGENOTROPHIC METHANOGEN MEDIUM
| NH4Cl | 0.54 | g |
| KH2PO4 | 0.14 | g |
| MgCl2·6H2O | 3.0 | g |
| CaCl2·2H2O | 0.15 | g |
| NaCl | 20.0 | g |
| FeCl2 solution (see Medium No. 187) | 1.0 | ml |
| Trace element solution (see Medium No. 187) | 1.0 | ml |
| Selenite-tungstate solution (see Medium No. 431) | 1.0 | ml |
| Sodium acetate·3H2O | 0.27 | g |
| NaHCO3 | 2.5 | g |
| Resazurin | 1.0 | mg |
| Distilled water | 1.0 | L |
Mix components, 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), seal with butyl rubber stoppers and autoclave. Aseptically and anaerobically add per liter the following solutions (autoclaved or filter-sterilized, and stored under a N2 atmosphere):
| Trace vitamins* (see Medium No. 197) | 2.0 | ml |
| 0.5 M Coenzyme M solution | 10.0 | ml |
| Titanium citrate solution* (see below) | 10.0 | ml |
Titanium citrate solution:
| Titanium(III) chloride | 0.75 | g |
| Sodium citrate·2H2O | 2.94 | g |
| Distilled water | 55.0 | ml |
Mix components and centrifuge at 8,000 xg for 10 min. Filter-sterilize the supernatant and store under a N2 atmosphere.
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