Main sol. 161
Internal Solution-ID: S277
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Clarified rumen fluid | 300.00 | ml | - | - | |
K2HPO4 | 0.30 | g | 0.3 | 1.721 | |
KH2PO4 | 0.30 | g | 0.3 | 2.202 | |
(NH4)2SO4 | 0.30 | g | 0.3 | 2.268 | |
NaCl | 0.60 | g | 0.599 | 10.257 | |
MgSO4 x 7 H2O | 0.13 | g | 0.13 | 0.527 | |
CaCl2 x 2 H2O
(0.1% w/v) |
8.00 | ml | 0.008 | 0.054 | |
Modified Wolin's mineral solution | 10.00 | ml | - | - | |
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) |
2.00 | ml | 0.002 | 0.007 | |
Yeast extract
(DIFCO) |
1.00 | g | 0.999 | - | |
Trypticase
(BBL) |
1.00 | g | 0.999 | - | |
Fatty acid mixture | 20.00 | ml | - | - | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
NaHCO3 | 2.00 | g | 1.998 | 23.784 | |
Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
L-Cysteine HCl x H2O | 0.50 | g | 0.5 | 2.844 | |
Na2S x 9 H2O | 0.50 | g | 0.5 | 2.08 | |
Distilled water | 660.00 | ml | - | - | |
1 Dissolve ingredients (except bicarbonate, vitamins, cysteine and sulfide) and sparge medium for 30 - 45 min with 80% H2 and 20% CO2 gas mixture to make it anoxic. Add and dissolve bicarbonate and adjust pH to 6.8, then distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add cysteine and sulfide from sterile anoxic stock solutions autoclaved under 100% N2 gas atmosphere. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of complete medium to 6.5. For incubation use sterile 80% H2 and 20% CO2 gas mixture at two atmospheres of pressure. |
161 | METHANOFOLLIS MEDIUM |