Main sol. 825a
Internal Solution-ID: S1674
Equipment needed: Hungate tubes
Filter
Autoclave
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NH4Cl | 1.0 | g | 0.979 | 18.311 | |
| K2HPO4 | 0.3 | g | 0.294 | 1.687 | |
| KH2PO4 | 0.3 | g | 0.294 | 2.159 | |
| MgCl2 x 6 H2O | 0.2 | g | 0.196 | 0.964 | |
| CaCl2 x 2 H2O | 0.1 | g | 0.098 | 0.666 | |
| KCl | 0.1 | g | 0.098 | 1.314 | |
| NaCl | 0.6 | g | 0.588 | 10.056 | |
| Modified Wolin's mineral solution | 10.0 | ml | - | - | |
| Na-acetate | 0.5 | g | 0.49 | 5.97 | |
| Yeast extract | 1.0 | g | 0.979 | - | |
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | 4.9e-4 | 0.002 | |
| NaHCO3 | 4.0 | g | 3.918 | 46.636 | |
|
Methanol
(15% v/v) |
10.0 | ml | 7.757 | 242.092 | |
| Wolin's vitamin solution (10x) | 1.0 | ml | - | - | |
| L-Cysteine HCl x H2O | 0.5 | g | 0.49 | 2.788 | |
| Na2S x 9 H2O | 0.5 | g | 0.49 | 2.039 | |
| Distilled water | 1000.0 | ml | - | - | |
| 1 Dissolve ingredients except bicarbonate, methanol, vitamins, cysteine and sulfide. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, adjust pH to 7.0, then dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add methanol (15% v/v solution), vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 6.8 - 7.0, if necessary. | |||||
Media with this solution
| 825a | METHANOBACTERIUM II MEDIUM (N2/CO2) |