Medium data
Search for medium no. = [228].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
228 METHANOCOCCUS McC MEDIUM
| General salts solution (see below) | 500.0 | ml |
| Trace minerals solution (see below) | 10.0 | ml |
| Iron stock solution (see below) | 5.0 | ml |
| NaCl | 22.0 | g |
| K2HPO4 | 0.14 | g |
| Sodium acetate | 1.36 | g |
| Yeast extract (BD-Difco) | 2.0 | g |
| Resazurin | 1.0 | mg |
| NaHCO3 | 5.0 | g |
| L-Cysteine·HCl·H2O | 0.5 | g |
| Na2S·9H2O | 0.5 | g |
| Distilled water | 485.0 | ml |
Mix ingredients, except cysteine·HCl and Na2S·9H2O, and bring to a boil for 5-10 sec. Cool down under a stream of N2-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), seal with butyl rubber stoppers, and autoclave. 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 vessels to 200 kPa H2-CO2 (80:20).
General salts solution:
| MgCl2·6H2O | 5.5 | g |
| MgSO4·7H2O | 6.9 | g |
| NH4Cl | 1.0 | g |
| KCl | 0.67 | g |
| CaCl2·2H2O | 0.28 | g |
Trace minerals solution:
| Nitrilotriacetic acid | 1.5 | g |
| Fe(NH4)2(SO4)2·6H2O | 0.2 | g |
| Na2SeO3 | 0.1 | g |
| Na2MoO4·2H2O | 0.1 | g |
| Na2WO4·2H2O | 1.0 | g |
| MnSO4·xH2O | 0.1 | g |
| ZnSO4·7H2O | 0.1 | g |
| NiCl2·6H2O | 0.025 | g |
| CuSO4·5H2O | 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.
Iron stock solution:
Dissolve 0.2 g of Fe(NH4)2(SO4)2·6H2O in 5 ml of distilled water containing 2 drops of concentrated HCl. When the Fe(NH4)2(SO4)2·6H2O crystal have dissolved, bring volume up to 100 ml.
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