Main sol. J231
Internal Solution-ID: S3908
Equipment needed: Autoclave
Anaerobic workbench
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| KH2PO4 | 0.30 | g | 0.3 | 2.204 | |
| (NH4)2SO4 | 1.50 | g | 1.5 | 11.352 | |
| NaCl | 0.60 | g | 0.6 | 10.267 | |
| MgSO4 x 7 H2O | 0.12 | g | 0.12 | 0.487 | |
| CaCl2 x 2 H2O | 0.08 | g | 0.08 | 0.544 | |
| FeSO4 x 7 H2O | 4.00 | mg | 0.004 | 0.014 | |
| K2HPO4 | 0.15 | g | 0.15 | 0.861 | |
| Na2CO3 | 4.00 | g | 4 | 37.74 | |
| Trace vitamins | 10.00 | ml | - | - | |
| Trace element solution | 10.00 | ml | - | - | |
| L-Cysteine HCl x H2O | 0.50 | g | 0.5 | - | |
| Na2S x 9 H2O | 0.50 | g | 0.5 | 2.082 | |
| Resazurin | 1.00 | mg | 1.0e-3 | 0.004 | |
| Distilled water | 980.00 | ml | - | - | |
| 1 Mix ingredients, except cysteine x HCl and Na2S x 9H2O, and adjust pH to 7.2. Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20). Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Separately autoclave cysteine x HCl and Na2S x 9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the cysteine x HCl and Na2S x 9H2O solutions. Pressurize the inoculated bottles to 100 kPa H2-CO2 (80:20). | |||||
Media with this solution
| J231 | METHANOBACTERIUM THERMOAUTOTROPHICUM MEDIUM |