Main sol. 332
Internal Solution-ID: S610
Equipment needed: Autoclave
Hungate tubes
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NH4Cl | 1.0 | g | 1 | 18.695 | |
| K2HPO4 x 3 H2O | 0.4 | g | 0.4 | 2.297 | |
| MgCl2 x 6 H2O | 0.1 | g | 0.1 | 0.492 | |
| Na-formate | 5.0 | g | 5 | 73.522 | |
| Na-acetate | 1.0 | g | 1 | 12.19 | |
|
Trypticase peptone
(BD BBL) |
1.0 | g | 1 | - | |
|
Yeast extract
(OXOID) |
1.0 | g | 1 | - | |
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | 5.0e-4 | 0.002 | |
| L-Cysteine HCl x H2O | 0.5 | g | 0.5 | 2.847 | |
| Na2CO3 | 1.5 | g | 1.5 | 14.153 | |
| Na2S x 9 H2O | 0.2 | g | 0.2 | 0.833 | |
| Distilled water | 1000.0 | ml | - | - | |
| 1 Dissolve ingredients except cysteine, carbonate and sulfide. Sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve cysteine, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture and sulfide from a sterile anoxic stock solution prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 6.8 - 7.0, if necessary. | |||||
| 2 Note: After growth has started add sterile 80% H2 and 20% CO2 gas mixture to 0.5 - 1 bar overpressure. | |||||
Media with this solution
| 332 | METHANOCULLEUS BOURGENSIS MEDIUM |