Equipment needed: Autoclave
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NaCl | 20.00 | g | 20 | 342.231 | |
| MgSO4 x 7 H2O | 7.00 | g | 7 | 28.4 | |
| MgCl2 x 6 H2O | 5.50 | g | 5.5 | 27.053 | |
| KCl | 0.65 | g | 0.65 | 8.719 | |
| NaBr | 0.10 | g | 0.1 | 0.972 | |
| NH4Cl | 0.30 | g | 0.3 | 5.609 | |
| K2HPO4 | 0.20 | g | 0.2 | 1.148 | |
| CaCl2 x 2 H2O | 0.50 | g | 0.5 | 3.401 | |
| Modified Wolin's mineral solution II | 10.00 | ml | - | - | |
| Yeast extract | 0.20 | g | 0.2 | - | |
|
Sulfur
(powdered) |
5.00 | g | 5 | 155.909 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Adjust pH to 7.0 using H2SO4. | |||||
| 2 Prepare the medium under H2 gas and fill 20 ml medium into 100 ml serum bottles and seal with a rubber stopper. Steam/heat the medium in an almost boiling waterbath for 3 hours on each of 3 successive days to sterilize. | |||||
| 3 Add bicarbonate from a autoclaved and neutralized stock solution. | |||||
| NaHCO3 | 2.00 | g/l | 2 | 23.808 | |
| 4 Check pH and adjust accordingly. | |||||
| 5 Give an atmosphere of H2/CO2/O2 (78/20/2) with an overpressure of two atmospheres. | |||||
Media with this solution
| 889 | ISS1 MEDIUM |