Main sol. J524
Internal Solution-ID: S4319
Equipment needed: Autoclave
Filter
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NaCl | 30.00 | g | 29.528 | 505.263 | |
| KH2PO4 | 0.14 | g | 0.138 | 1.013 | |
| CaCl2 x 2 H2O | 0.80 | g | 0.787 | 5.356 | |
| MgSO4 x 7 H2O | 3.40 | g | 3.346 | 13.577 | |
| MgCl2 x 6 H2O | 4.20 | g | 4.134 | 20.334 | |
| KCl | 0.33 | g | 0.325 | 4.357 | |
| NaNO3 | 0.25 | g | 0.246 | 2.895 | |
| NH4Cl | 0.25 | g | 0.246 | 4.6 | |
| Na2SiO3 x 9 H2O | 0.20 | g | 0.197 | - | |
| Fe2(SO4)3 x n H2O | 20.00 | mg | 0.02 | - | |
| Na2SeO3 x 5 H2O | 0.50 | mg | 4.9e-4 | 0.002 | |
| NiCl2 x 6 H2O | 0.50 | mg | 4.9e-4 | 0.004 | |
| Trace mineral solution | 3.00 | ml | - | - | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Mix components, adjust pH to 6.0 - 6.5 and autoclave. After cooling, add the following solutions (filter-sterilized) to the medium: | |||||
| Trace vitamins | 1.00 | ml | - | - | |
|
NaHCO3
(5%) |
10.00 | ml | - | - | |
|
CuSO4
(1 mM) |
2.00 | ml | - | - | |
| 2 Distribute the medium into culture vessels (leave >80% volume gas phase) under a N2-CO2 (4:1) gas mixture and seal with butyl rubber stoppers. Pressurize the culture vessels to 200 kPa with methane gas. Finally add O2 gas to make up 2% O2 concentration in the gas phase. Readjust pH to 6.0 - 6.3, if necessary. | |||||
Media with this solution
| J524 | METHANE OXIDIZING BACTERIUM MEDIUM |