Main sol. 704
Internal Solution-ID: S1455
Equipment needed: Centrifuge
Autoclave
Hungate tubes
Filter
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Mineral solution | 75.00 | ml | - | - | |
| Clarified rumen fluid | 150.00 | ml | - | - | |
| K2HPO4 | 0.30 | g | 0.3 | 1.721 | |
|
Trypticase peptone
(BD BBL) |
2.00 | g | 1.998 | - | |
|
Yeast extract
(OXOID) |
2.00 | g | 1.998 | - | |
| Volatile fatty acid mixture | 3.10 | ml | - | - | |
|
Haemin solution
(0.05% w/v) |
2.00 | ml | - | - | |
| Glycerol | 0.50 | g | 0.5 | 5.424 | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
| Na2CO3 | 4.00 | g | 3.996 | 37.702 | |
| D-Glucose | 1.00 | g | 0.999 | 5.545 | |
| Maltose | 1.00 | g | 0.999 | 2.919 | |
| Cellobiose | 1.00 | g | 0.999 | 2.919 | |
|
Starch
(soluble) |
1.00 | g | 0.999 | - | |
| L-Cysteine HCl x H2O | 0.25 | g | 0.25 | 1.422 | |
| Na2S x 9 H2O | 0.25 | g | 0.25 | 1.04 | |
| Distilled water | 770.00 | ml | - | - | |
| 1 Dissolve ingredients (except carbonate, carbohydrates, cysteine and sulfide) and sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add the carbonate and equilibrate the medium with the CO2 gas to pH 6.8. Distribute under 100% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Thereafter, add glucose, maltose, cellobiose, starch, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Cellobiose should be sterilized by filtration. Adjust pH of complete medium to 6.7 - 6.8, if necessary. | |||||
Media with this solution
| 704 | BUTYRIVIBRIO SP. MEDIUM |