| | (NH4)2SO4 | 1.3 | g | | | KH2PO4 | 0.28 | g | | | MgSO4·7H2O | 0.25 | g | | | CaCl2·2H2O | 0.07 | g | | | FeCl3·6H2O | 0.02 | g | | | MnCl2·4H2O | 1.8 | mg | | | Na2B4O7·10H2O | 4.5 | mg | | | ZnSO4·7H2O | 0.22 | mg | | | CuCl2·2H2O | 0.05 | mg | | | Na2MoO4·2H2O | 0.03 | mg | | | VOSO4·xH2O | 0.03 | mg | | | CoSO4·7H2O | 0.01 | mg | | | Bacto Yeast Extract (Difco) | 0.5 | g | | | Sulfur (powder) | 1 | g | | | Resazurin | 1 | mg | | | Na2S·9H2O | 0.5 | g | | | Distilled water | 1 | L | | | | | Mix ingredients except for Bacto Yeast Extract, sulfur and Na2S·9H2O, and adjust pH to 2.0 with H2SO4. Dispense the medium into suitable culture vessels under a stream of H2/CO2 (80/20) and seal with butyl rubber stoppers. Separately autoclave Bacto Yeast Extract and Na2S·9H2O as 5% solutions under a N2 atmosphere. Sulfur is sterilized separately by autoclaving for 60 min at 105°C on each of successive days. Prior to inoculation, add Bacto Yeast Extract, sulfur and Na2S·9H2O, and pressurize the inoculated vessels to 150 kPa with H2/CO2. |
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