Main sol. 734
Internal Solution-ID: S1504
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Clarified rumen fluid | 400.00 | ml | - | - | |
NaCl | 1.00 | g | 0.997 | 17.06 | |
KCl | 0.50 | g | 0.499 | 6.687 | |
MgCl2 x 6 H2O | 0.40 | g | 0.399 | 1.962 | |
CaCl2 x 2 H2O | 0.10 | g | 0.1 | 0.678 | |
NH4Cl | 0.30 | g | 0.299 | 5.592 | |
KH2PO4 | 0.20 | g | 0.199 | 1.465 | |
Na2SO4 | 0.15 | g | 0.15 | 1.053 | |
Casamino acids
(BD Bacto) |
0.50 | g | 0.499 | - | |
Yeast extract
(OXOID) |
0.50 | g | 0.499 | - | |
Nutrient broth
(BD Difco) |
2.00 | g | 1.994 | - | |
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 | 0.997 | 9.407 | |
MOPS
(SIGMA) |
2.10 | g | 2.094 | 10.005 | |
Seven vitamins solution | 1.00 | ml | - | - | |
DL-Dithiothreitol
(DTT) |
0.16 | g | 0.16 | 1.034 | |
Distilled water | 600.00 | ml | - | - | |
1 Dissolve ingredients (except carbonate, MOPS buffer, vitamins and DTT), bring medium to the boil, then cool to room temperature under 80% H2 and 20% CO2 gas mixture. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add MOPS buffer adjusted to pH 7.2, vitamins and DTT from anoxic stock solutions prepared under 100% N2 gas and sterilized by filtration and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 7.2, if necessary. |
734 | METHANOBREVIBACTER CURVATUS MEDIUM |