Main sol. 1318
Internal Solution-ID: S2639
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
MgCl2 x 6 H2O | 0.40 | g | 0.398 | 1.96 | |
CaCl2 x 2 H2O | 0.10 | g | 0.1 | 0.677 | |
NH4Cl | 0.10 | g | 0.1 | 1.862 | |
KH2PO4 | 0.20 | g | 0.199 | 1.464 | |
KCl | 0.50 | g | 0.498 | 6.68 | |
Yeast extract | 0.20 | g | 0.199 | - | |
Na-acetate | 0.09 | g | 0.09 | 1.093 | |
Trace element solution SL-10 | 1.00 | ml | - | - | |
Selenite-tungstate solution | 1.00 | ml | - | - | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
Na2CO3 | 1.00 | g | 0.996 | 9.397 | |
Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
Seven vitamins solution | 1.00 | ml | - | - | |
L-Cysteine HCl x H2O | 0.30 | g | 0.299 | 1.701 | |
DL-Dithiothreitol | 0.30 | g | 0.299 | 1.937 | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients (except carbonate, vitamins, cysteine, and DTT) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add vitamins (sterilized by filtration), cysteine, and DTT from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. The pH of the complete medium should be 7.0. | |||||
2 After inoculation pressurize vials to 1 bar overpressure with sterile 100% H2 gas. |
1318 | METHANOCELLA CONRADII MEDIUM |