| |  | NaCl | 13.85 | g |  |  | MgSO4·7H2O | 3.5 | g |  |  | MgCl2·6H2O | 2.75 | g |  |  | KCl | 0.325 | g |  |  | NaBr | 0.05 | g |  |  | H3BO3 | 15 | mg |  |  | SrCl2·6H2O | 7.5 | mg |  |  | KI | 0.05 | mg |  |  | CaCl2 | 0.75 | g |  |  | KH2PO4 | 0.5 | g |  |  | (NH4)2Ni(SO4)2·6H2O | 2 | mg |  |  | Trace minerals* | 10 | ml |  |  | Bacto Yeast Extract (Difco) | 2 | g |  |  | Citric acid | 5 | mg |  |  | Sulfur (powder) | 30 | g |  |  | Resazurin | 1 | mg |  |  | Na2S·9H2O | 0.3 | g |  |  | Distilled water | 1 | L |  |  |  |  |  | Mix ingredients except sulfur and Na2S·9H2O, adjust pH to 5.5 with H2SO4, and autoclave under H2/CO2 (80/20) conditions. Separately autoclave neutralize Na2S·9H2O solution. Sulfur is sterilized separately by autoclaving for 60 min at 105℃ on each of 3 successive days. Aseptically and anaerobically add sterile sulfur to the medium under a H2/CO2 (80/20) atmosphere. Prior to inoculation, add Na2S·9H2O solution, and readjust pH to 5.5 if necessary. Pressurize inoculated bottles to 200 kPa with H2/CO2 (80/20). |  |  |  |  |  |  |  |  |  |  |  | *Trace minerals |  |  |  |  | Nitrilotriacetic acid | 1.5 | g |  |  |  |  | MgSO4·7H2O | 3 | g |  |  |  |  | MnSO4·xH2O | 0.5 | g |  |  |  |  | NaCl | 1 | g |  |  |  |  | FeSO4·7H2O | 0.1 | g |  |  |  |  | CoSO4·7H20 | 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 |  |  |  |  | Distilled water | 1 | L |  |  |  |  |  | Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0. | 
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