Main sol. 339a
Internal Solution-ID: S5601
Equipment needed: Filter
Autoclave
Hungate tubes
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
|
dehydrated Wilkins-Chalgren medium
(Oxoid CM0643) |
33.0 | g | 33 | - | |
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | 5.0e-4 | - | |
| L-Cysteine HCl | 0.3 | g | 0.3 | 1.903 | |
| D-Glucose | 5.0 | g | 5 | 27.753 | |
| Na2CO3 | 1.0 | g | - | - | |
| Distilled water | 1000.0 | ml | - | - | |
| 1 Dissolve 33.0 g/l dehydrated Wilkins-Chalgren medium (Oxoid CM0643) in distilled water and add 0.5 ml/l Na-resazurin solution (0.1% w/v), bring to the boil and cool to room temperature while sparging with 80% N2 and 20% CO2 gas mixture. Add L-cysteine-HCL x H2O (0.3 g/l), dispense the medium into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving supplement medium with 5.0 g/l D-glucose added from a sterile anoxic stock solution sterilized by filtration. Adjust pH of medium to 6.8 with a sterile anoxic stock solution of Na2CO3 (5% w/v) prepared under 80% N2 and 20% CO2 gas atmosphere. | |||||
| 2 Note: In case resazurin is not colourless addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) can be used to completely reduce the medium. | |||||
Media with this solution
| 339a | WILKINS-CHALGREN ANAEROBE BROTH (N2/CO2) |