Main sol. J324
Internal Solution-ID: S4041
Equipment needed: Autoclave
Anaerobic workbench
Filter
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NaCl | 30.00 | g | 29.412 | 503.281 | |
| K2HPO4 | 0.14 | g | 0.137 | 0.788 | |
| CaCl2 x 2 H2O | 1.00 | g | 0.98 | 6.669 | |
| NaNO3 | 0.80 | g | 0.784 | 9.228 | |
| MgSO4 x 7 H2O | 3.40 | g | 3.333 | 13.524 | |
| MgCl2 x 6 H2O | 4.18 | g | 4.098 | 20.157 | |
| KCl | 0.33 | g | 0.324 | 4.34 | |
| NiCl2 x 6 H2O | 5.00 | mg | 0.005 | 0.038 | |
| Na2SeO3 | 1.00 | mg | 9.8e-4 | - | |
| Fe2(SO4)3 x n H2O | 0.05 | g | 0.049 | - | |
| H2WO4 | 5.00 | mg | 0.005 | 0.02 | |
| Trace mineral solution | 10.00 | ml | - | - | |
| Trace vitamins | 10.00 | ml | - | - | |
| NaHCO3 | 1.00 | g | 0.98 | 11.67 | |
| Yeast extract | 1.00 | g | 0.98 | - | |
| Sodium acetate | 0.40 | g | 0.392 | 4.781 | |
|
Sulfur
(powder) |
30.00 | g | 29.412 | 917.111 | |
| Na2S x 9 H2O | 0.50 | g | 0.49 | 2.041 | |
| Resazurin | 1.00 | mg | 9.8e-4 | 0.004 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Mix ingredients except Trace vitamins, NaHCO3, sulfur and Na2S x 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 x 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). | |||||
Media with this solution
| J324 | MMJY MEDIUM |