Main sol. 1718
Internal Solution-ID: S3555
Equipment needed: Autoclave
Hungate tubes
Filter
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| KH2PO4 | 0.20 | g | 0.199 | 1.465 | |
| NH4Cl | 0.25 | g | 0.249 | 4.66 | |
| NaCl | 1.00 | g | 0.997 | 17.06 | |
| MgSO4 x 7 H2O | 0.40 | g | 0.399 | 1.618 | |
| KCl | 0.50 | g | 0.499 | 6.687 | |
| CaCl2 x 2 H2O | 0.15 | g | 0.15 | 1.017 | |
| Trace element solution SL-10 | 1.00 | ml | - | - | |
| Selenite-tungstate solution | 1.00 | ml | - | - | |
| Na-acetate | 0.80 | g | 0.798 | 9.723 | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
| Na2CO3 | 1.00 | g | 0.997 | 9.407 | |
| Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
| Na2S x 9 H2O | 0.50 | g | 0.499 | 2.076 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients (except carbonate, vitamins and sulfide) and sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Then dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas (vitamins are sterilized by filtration) and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of the complete medium to 7.0 - 7.4, if necessary. | |||||
| 2 After inoculation, pressurize culture vessels with sterile 80% H2 and 20% CO2 gas mixture to 1 bar overpressure. | |||||
Media with this solution
| 1718 | METHANOSPIRILLUM F.2.7.3 MEDIUM |