Main sol. J251
Internal Solution-ID: S3935
Equipment needed: Autoclave
Anaerobic workbench
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Mineral solution 1 | 37.5 | ml | - | - | |
| Mineral solution 2 | 37.5 | ml | - | - | |
| (NH4)2Ni(SO4)2 x 6 H2O | 3.0 | mg | 0.003 | 0.007 | |
| FeSO4 x 7 H2O | 2.0 | mg | 0.002 | 0.007 | |
|
Yeast extract
(BD-Difco) |
2.0 | g | 1.97 | - | |
|
Trypticase peptone
(BD-BBL) |
2.0 | g | 1.97 | - | |
| Trace vitamins | 10.0 | ml | - | - | |
| Trace minerals | 10.0 | ml | - | - | |
| Na2SO4 | 3.4 | g | 3.35 | 23.583 | |
| NaHCO3 | 2.0 | g | 1.97 | 23.456 | |
| Resazurin | 1.0 | mg | 9.9e-4 | 0.004 | |
| Na2S x 9 H2O | 0.5 | g | 0.493 | 2.051 | |
| L-Cysteine HCl x H2O | 0.5 | g | 0.493 | - | |
| Distilled water | 920.0 | ml | - | - | |
| 1 Mix ingredients, except NaHCO3, L-cysteine x HCl x H2O and Na2S x 9H2O, and bring to a boil for 5-10 sec. Cool down under a stream of H2-CO2 (80:20), and add NaHCO3 to the medium. Dispense the medium into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20) gas mixture and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight and adjust pH to 6.5, if necessary. Separately autoclave L-cysteine x HCl and Na2S x 9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the L-cysteine x HCl and Na2S x 9H2O solutions. Pressurize the inoculated bottles to 200 kPa H2-CO2 (80:20). | |||||
Media with this solution
| J251 | METHANOTHERMUS FERVIDUS MEDIUM |