Main sol. 991
Internal Solution-ID: S2020
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
(NH4)2SO4 | 1.30 | g | 1.286 | 9.731 | |
KH2PO4 | 0.28 | g | 0.277 | 2.035 | |
MgSO4 x 7 H2O | 0.25 | g | 0.247 | 1.003 | |
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 | - | - | |
Trisodium citrate x 2 H2O | 2.94 | g | 2.908 | 9.888 | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 4.9e-4 | 0.002 | |
Sulfur
(powdered) |
10.00 | g | 9.891 | 308.425 | |
Yeast extract
(OXOID) |
0.50 | g | 0.495 | - | |
Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
Na2S x 9 H2O | 0.50 | g | 0.495 | 2.059 | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients (except sulfur, yeast extract, vitamins and sulfide), adjust pH to 3.5 with 10 N H2SO4 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 Hungate-type tubes or serum vials (e.g., 20 ml medium in 100 ml bottles) containing already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a water bath to 90 - 100°C for 1 - 2 hours on each of 3 successive days. Add yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 4.0 - 4.5, if necessary. | |||||
2 After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. |
991 | CALDISPHAERA MEDIUM |