Medium data
Search for medium no. = [324].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
324 MMJY MEDIUM
| NaCl | 30.0 | g |
| K2HPO4 | 0.14 | g |
| CaCl2·2H2O | 1.0 | g |
| NaNO3 | 0.8 | g |
| MgSO4·7H2O | 3.4 | g |
| MgCl2·6H2O | 4.18 | g |
| KCl | 0.33 | g |
| NiCl2·6H2O | 5.0 | mg |
| Na2SeO3 | 1.0 | mg |
| Fe2(SO4)3·xH2O | 0.05 | g |
| H2WO4 | 5.0 | mg |
| Trace mineral solution (see below) | 10.0 | ml |
| Trace vitamins (see Medium No. 197) | 10.0 | ml |
| NaHCO3 | 1.0 | g |
| Yeast extract | 1.0 | g |
| Sodium acetate | 0.4 | g |
| Sulfur (powder) | 30.0 | g |
| Na2S·9H2O | 0.5 | g |
| Resazurin | 1.0 | mg |
| Distilled water | 1.0 | L |
Mix ingredients except Trace vitamins, NaHCO3, sulfur and Na2S·9H2O, adjust pH to 7.5 - 8.0 and autoclave. Filter-sterilize Trace vitamins and 5% NaHCO3 solution and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Separately autoclave 5% Na2S·9H2O solution under a N2 atmosphere. Distribute the medium into culture vessels containing appropriate amount of sulfur (e.g., 20 ml of medium in 120 ml serum bottles containing 0.6 g sulfur) under a stream of H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Aseptically and anaerobically add the sterile Na2S solution. Pressurize the inoculated bottle to 200 kPa H2-CO2 (4:1, v/v).
Trace mineral solution:
| Nitrilotriacetic acid | 1.5 | g |
| MnSO4·xH2O | 0.5 | g |
| CoSO4·7H2O | 0.5 | 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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