Main sol. 358a
Internal Solution-ID: S698
Equipment needed: Serum bottle
Gassing station
Water bath
Autoclave
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| (NH4)2SO4 | 1.30 | g | 1.287 | 9.741 | |
| KH2PO4 | 0.28 | g | 0.277 | 2.037 | |
| MgSO4 x 7 H2O | 0.25 | g | 0.248 | 1.004 | |
| CaCl2 x 2 H2O | 0.07 | g | 0.069 | 0.471 | |
| FeCl3 x 6 H2O | 0.02 | g | 0.02 | 0.073 | |
| Allen's trace element solution | 10.00 | ml | - | - | |
|
Sulfur
(powder) |
5.00 | g | 4.951 | 154.365 | |
|
Yeast extract
(OXOID) |
0.50 | g | 0.495 | - | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients, except sulfur and yeast extract, bring medium to the boil, then cool to room temperature under 80% H2 and 20% CO2 gas mixture and adjust pH to 2.5 using 10 N H2SO4. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 100 ml serum bottles) containing already the appropriate amount of sulfur. For sterilization sealed bottles with medium are heated in a boiling water bath for 2 - 3 h on each of 3 successive days. Add yeast extract from a sterile anoxic stock solution prepared under 100% N2 gas atmosphere. | |||||
| 2 Pressurize inoculated bottles to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. | |||||
| 3 Note: Inoculate with 5% (w/v) culture. Incubate without shaking. | |||||
Media with this solution
| 358a | ACIDIANUS MEDIUM (ANAEROBIC) |