Main sol. 814
Internal Solution-ID: S1653
Equipment needed: Autoclave
Hungate tubes
Filter
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NH4Cl | 0.40 | g | 0.4 | 7.478 | |
| MgCl2 x 6 H2O | 0.03 | g | 0.03 | 0.148 | |
| NaCl | 0.45 | g | 0.45 | 7.7 | |
| K2HPO4 | 0.50 | g | 0.5 | 2.871 | |
|
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) |
3.00 | ml | 0.003 | 0.011 | |
| Modified Wolin's mineral solution | 10.00 | ml | - | - | |
|
Tryptone
(BD Bacto) |
1.00 | g | 1 | 14.069 | |
| Na-formate | 3.20 | g | 3.2 | 47.054 | |
| Na-acetate | 1.00 | g | 1 | 12.19 | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - | |
| Na2CO3 | 1.00 | g | 1 | 9.435 | |
| Seven vitamins solution | 1.00 | ml | - | - | |
| L-Cysteine HCl x H2O | 0.25 | g | 0.25 | 1.423 | |
| Na2S x 9 H2O | 0.25 | g | 0.25 | 1.041 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients except carbonate, vitamins, cysteine and sulfide, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 8.0 - 8.5. | |||||
Media with this solution
| 814 | METHANOBACTERIUM SUBTERRANEUM MEDIUM |