339a .
WILKINS-CHALGREN ANAEROBE BROTH (N2/CO2)
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Selected strain: Succinatimonas hippei DSM 22608
This strain uses the standard medium without modifications.
Growth conditions: anaerobic
37 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
6.8
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 | ||
|
|
D-Glucose | 5.0 | g | ||
|
|
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. | |||||