Main sol. 119
Internal Solution-ID: S207
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
KH2PO4 | 0.50 | g | 0.499 | 3.663 | |
MgSO4 x 7 H2O | 0.40 | g | 0.399 | 1.618 | |
NaCl | 0.40 | g | 0.399 | 6.824 | |
NH4Cl | 0.40 | g | 0.399 | 7.456 | |
CaCl2 x 2 H2O | 0.05 | g | 0.05 | 0.339 | |
Trace element solution SL-10 | 1.00 | ml | - | - | |
Yeast extract
(OXOID) |
1.00 | g | 0.997 | - | |
Na-acetate | 1.00 | g | 0.997 | 12.154 | |
Na-formate | 2.00 | g | 1.994 | 29.321 | |
FeSO4 x 7 H2O solution (0.1% w/v) | 2.00 | ml | - | - | |
Sludge fluid | 50.00 | ml | - | - | |
Fatty acid mixture | 20.00 | ml | - | - | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
NaHCO3 | 4.00 | g | 3.988 | 47.473 | |
L-Cysteine HCl x H2O | 0.50 | g | 0.499 | 2.838 | |
Na2S x 9 H2O | 0.50 | g | 0.499 | 2.076 | |
Distilled water | 930.00 | ml | - | - | |
1 Dissolve ingredients except bicarbonate, cysteine and sulfide. Sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, adjust pH to 6.5 and dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 6.8 - 7.0, if necessary. | |||||
2 Note: After growth has started and the culture is becoming turbid add sterile 80% H2 and 20% CO2 gas mixture to 0.5 - 1 bar overpressure. |
119 | METHANOBACTERIUM MEDIUM |