Main sol. 1455
Internal Solution-ID: S2989
Equipment needed: Filter
Autoclave
Hungate tubes
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| K2HPO4 | 0.23 | g | 0.229 | 1.313 | |
| KH2PO4 | 0.23 | g | 0.229 | 1.68 | |
| NaCl | 0.45 | g | 0.447 | 7.654 | |
| (NH4)2SO4 | 0.45 | g | 0.447 | 3.385 | |
| CaCl2 x 2 H2O | 0.06 | g | 0.06 | 0.406 | |
| MgSO4 x 7 H2O | 0.09 | g | 0.089 | 0.363 | |
|
Brain heart infusion
(BD BBL) |
6.00 | g | 5.964 | - | |
| Yeast extract | 5.00 | g | 4.97 | - | |
| Volatile fatty acid mixture | 3.10 | ml | - | - | |
|
Haemin solution
(0.05% w/v) |
2.00 | ml | - | - | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
| NaHCO3 | 6.40 | g | 6.362 | 75.73 | |
| Disodium succinate | 5.00 | g | 4.97 | 30.671 | |
| Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
| L-Cysteine HCl x H2O | 0.30 | g | 0.298 | 1.698 | |
| Na2S x 9 H2O | 0.30 | g | 0.298 | 1.242 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients except bicarbonate, succinate, vitamins, cysteine and sulfide, then sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add the bicarbonate and equilibrate the medium with the CO2 gas to reach a pH between 6.5 and 7.0. Dispense medium under 100% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add succinate, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The vitamin solution should be sterilized by filtration. Prior to use adjust pH of complete medium to 6.6 - 6.8, if necessary. | |||||
Media with this solution
| 1455 | ALTERNATIVE RUMEN BACTERIA MEDIUM |