Medium data
Search for medium no. = [523].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
523 METHYLOTHERMUS SUBTERRANEUS MEDIUM
| NaCl | 1.0 | g |
| KH2PO4 | 0.014 | g |
| CaCl2·2H2O | 0.08 | g |
| MgSO4·7H2O | 0.34 | g |
| MgCl2·6H2O | 0.42 | g |
| KCl | 0.033 | g |
| NH4Cl | 0.5 | g |
| Na2SiO3·9H2O | 0.5 | g |
| Fe2(SO4)3·xH2O | 20.0 | mg |
| Na2SeO3·5H2O | 0.5 | mg |
| NiCl2·6H2O | 0.5 | mg |
| Trace mineral solution (see below) | 3.0 | ml |
| Distilled water | 1.0 | L |
Mix components thoroughly and autoclave. After cooling, add the following solutions (filter-sterilized) to the medium:
| Trace vitamins (see Medium No. 197) | 1.0 | ml |
| 8% NaHCO3 solution | 2.0 | ml |
| 1 mM CuSO4 solution | 1.0 | ml |
Then, distribute the medium into culture vessels (leave >80% gas phase, by volume) and seal with butyl rubber stoppers. Replace the gas phase with N2-CO2-CH4-O2 (40:15:40:5, by volume) and pressurize to 100 kPa with the same gas mixture. Check final pH of the medium to be around 6.0.
Trace mineral solution:
| Nitrilotriacetic acid | 1.5 | g |
| MnSO4·xH2O | 0.5 | g |
| CoSO4·7H2O | 0.18 | g |
| ZnSO4·7H2O | 0.18 | g |
| CuSO4·5H2O | 0.01 | g |
| AlK(SO4)2 | 0.02 | g |
| H3BO3 | 0.01 | g |
| Na2MoO4·2H2O | 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.
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