| | (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 | | | NaHCO3 | 0.85 | g | | | Bacto Yeast Extract (Difco) | 0.2 | g | | | Na2S2O4 | 25 | mg | | | Sulfur (powder) | 8 | g | | | Resazurin | 0.5 | mg | | | Distilled water | 1 | L | | | | | Mix ingredients except yeast extract, NaHCO3, Na2S2O4 and sulfur, and adjust pH to 6.5. Autoclave, then cool under a H2/CO2 (80/20) atmosphere. Separately autoclave yeast extract as a 10% solution under a N2 atmosphere. Filter-sterilize NaHCO3 as a 8% solution while gassing with N2. Sulfur is sterilized separately by autoclaving for 60 min at 105℃ on each of 3 successive days. Aseptically and anaerobically add the yeast extract and NaHCO3 solutions, and sulfur to the medium under a H2/CO2 (80/20) atmosphere. Readjust pH to 6.5 by further gassing with H2/CO2 (80/20) gas mixture, if necessary. Prior to inoculation, add filter-sterilized 2.5% Na2S2O4 solution. Pressurize inoculated bottles to 150 kPa with H2/CO2 (80/20). |
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