Medium data
Search for medium no. = [484].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
484 MODIFIED METHANOCALDOCOCCUS MEDIUM
| K2HPO4 | 0.14 | g |
| CaCl2·2H2O | 0.14 | g |
| NH4Cl | 0.25 | g |
| MgSO4·7H2O | 3.4 | g |
| MgCl2·6H2O | 2.7 | g |
| KCl | 0.33 | g |
| NaCl | 30.0 | g |
| Iron stock solution (see Medium No. 228) | 5.0 | ml |
| Trace minerals solution (see below) | 10.0 | ml |
| Trace vitamins (see Medium No. 197) | 10.0 | ml |
| Na2S2O3·5H2O | 1.0 | g |
| Yeast extract (BD-Difco) | 2.0 | g |
| 2-Mercaptoethanol | 0.4 | g |
| Resazurin | 1.0 | mg |
| Distilled water | 1.0 | L |
Mix ingredients except Na2S2O3·5H2O, yeast extract, Trace vitamins and 2-mercaptoethanol, and adjust pH to 6.0. Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20, v/v). Dispense the medium into suitable culture vessels under a stream of H2-CO2 (80:20, v/v), seal with butyl rubber stoppers and autoclave 115C for 40 min. Separately autoclave Na2S2O3·5H2O and 2-mercaptoethanol as 50 g/L solutions and yeast extract as 100 g/L solution under a N2 atmosphere. Filter-sterilize Trace vitamins and store under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the Na2S2O3·5H2O, yeast extract, Trace vitamins and 2-mercaptoethanol solutions to the medium. Pressurize the inoculated bottles 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 |
| NaCl | 1.0 | g |
| FeSO4·7H2O | 0.1 | g |
| CoSO4·7H2O | 0.1 | g |
| CaCl2·2H2O | 0.1 | g |
| ZnSO4·7H2O | 0.1 | g |
| CuSO4·5H2O | 0.01 | g |
| AlK(SO4)2 | 0.01 | g |
| H3BO3 | 0.01 | g |
| Na2MoO4·2H2O | 0.01 | g |
| NiCl2·6H2O | 0.025 | g |
| Na2SeO3·5H2O | 0.3 | mg |
| 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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