Main sol. 1092
Internal Solution-ID: S2199
Equipment needed: Filter
Autoclave
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
|
Sea Salt
(SIGMA) |
30.0 | g | 29.91 | - | |
| Wolfe's mineral elixir | 1.0 | ml | - | - | |
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | 5.0e-4 | 0.002 | |
|
Sulfur
(powdered) |
3.0 | g | 2.991 | 93.266 | |
| NaHCO3 | 2.5 | g | 2.493 | 29.67 | |
| Yeast extract | 2.0 | g | 1.994 | - | |
| Wolin's vitamin solution (10x) | 2.0 | ml | - | - | |
| Na2S x 9 H2O | 0.5 | g | 0.499 | 2.076 | |
| Distilled water | 1000.0 | ml | - | - | |
| 1 Dissolve ingredients (except sulfur, bicarbonate, yeast extract, vitamins and sulfide) and sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic serum vials (e.g., Balch-type tubes) containing already the appropriate amount of sulfur, only to 30% of their volume to allow for a large headspace. Autoclave at 110°C for 20 min. Add yeast extract, vitamins and sulfide form sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. The vitamin solution should be sterilized by filtration. Adjust pH of the complete medium to 7.0-7.2. | |||||
| 2 After inoculation pressurize vessels with sterile 80% H2 and 20% CO2 gas mixture to 2 bar overpressure. | |||||
Media with this solution
| 1092 | THERMOSULFIDIBACTER MEDIUM |