Main sol. 1104
Internal Solution-ID: S2220
Equipment needed: Hungate tubes
Water bath
Filter
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NaCl | 6.00 | g | 5.976 | 102.26 | |
| K2HPO4 | 0.50 | g | 0.498 | 2.859 | |
| NaHCO3 | 8.00 | g | 7.968 | 94.851 | |
| Na2CO3 | 22.00 | g | 21.912 | 206.744 | |
|
Sulfur
(powdered) |
1.50 | g | 1.494 | 46.586 | |
| NH4Cl | 0.20 | g | 0.199 | 3.724 | |
| MgCl2 x 6 H2O | 0.20 | g | 0.199 | 0.98 | |
| Trace elements solution (Pfennig & Lippert,1966) | 1.00 | ml | - | - | |
| Selenite-tungstate solution | 1.00 | ml | - | - | |
| Yeast extract | 0.05 | g | 0.05 | - | |
| Ethanol absolute | 0.60 | ml | - | - | |
| Na-acetate | 0.40 | g | 0.398 | 4.857 | |
| Na2S2O3 x 5 H2O | 2.50 | g | 2.49 | 10.033 | |
| Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
| Na2S x 9 H2O | 0.24 | g | 0.239 | 0.995 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve sodium chloride and hydrogenphosphate, then sparge solution with 100% N2 gas for 30 -45 min to make it anoxic. Add and dissolve carbonates, then dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials containing already the appropriate amount of sulfur, only to 30% of their volume to allow for a large headspace. Sterilize medium by heating cultivation vessels in a water bath to 90 - 100°C for 1- 2 hours on each of 3 successive days. After sterilization add ammonium chloride, magnesium chloride, trace elements, yeast extract, ethanol, acetate, thiosulfate, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The stock solutions of thiosulfate and vitamins should be sterilized by filtration. Adjust pH of the complete medium to 9.5. | |||||
| 2 After inoculation add sterile 100% H2 gas to 0.5 bar overpressure. | |||||
Media with this solution
| 1104 | DETHIOBACTER MEDIUM |