Main sol. 523
Internal Solution-ID: S1158
Equipment needed: Autoclave
Hungate tubes
Filter
Gassing station
Centrifuge
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Mineral solution | 38.00 | ml | - | - | |
| Clarified rumen fluid | 200.00 | ml | - | - | |
| K2HPO4 | 0.30 | g | 0.3 | 1.722 | |
|
Trypticase peptone
(BD BBL) |
2.00 | g | 2 | - | |
|
Yeast extract
(OXOID) |
0.50 | g | 0.5 | - | |
|
Haemin solution
(0.05% w/v) |
2.00 | ml | - | - | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - | |
| Na2CO3 | 4.00 | g | 4 | 37.74 | |
| Cellobiose | 5.00 | g | 5 | 14.607 | |
| L-Cysteine HCl x H2O | 0.25 | g | 0.25 | 1.423 | |
| Na2S x 9 H2O | 0.25 | g | 0.25 | 1.041 | |
| Distilled water | 760.00 | ml | - | - | |
| 1 Dissolve ingredients (except carbonate, cellobiose, cysteine and sulfide), bring medium to the boil, then cool to room temperature under 100% CO2 gas atmosphere. Add the carbonate and equilibrate the medium with the CO2 gas to pH 6.8. Distribute under CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add cellobiose, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Cellobiose has to be sterilized by filtration. Adjust pH of complete medium to 6.7 - 6.8, if necessary. | |||||
Media with this solution
| 523 | CLOSTRIDIUM CHARTATABIDUM MEDIUM |