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WILKINS-CHALGREN ANAEROBE BROTH
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* Medium for Bullifex porci DSM 105750
Add-on rumen fluid (50 ml/L) to stimulate growth; anaerobic,plus H2 (0,5 bar)
Growth conditions: 3-7 days
anaerobic
37 °C
pH 7.00
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
6.8 - 7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
dehydrated Wilkins-Chalgren medium
(Oxoid CM0643) |
33.0 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | ||
|
|
L-Cysteine HCl | 0.3 | g | ||
|
|
Rumen fluid | 50.0 | ml/l | ||
|
|
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 100% N2 gas. Add L-cysteine-HCL x H2O (0.3 g/l), dispense the medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving check pH and adjust to pH 6.8 - 7.0, if necessary. | |||||
| 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. | |||||