332 .
METHANOCULLEUS BOURGENSIS MEDIUM
What is this page?
Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final pH:
6.8 - 7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 1.0 | g | ||
|
|
K2HPO4 x 3 H2O | 0.4 | g | ||
|
|
MgCl2 x 6 H2O | 0.1 | g | ||
|
|
Na-formate | 5.0 | g | ||
|
|
Na-acetate | 1.0 | g | ||
|
|
Trypticase peptone
(BD BBL) |
1.0 | g | ||
|
|
Yeast extract
(OXOID) |
1.0 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | ||
|
|
L-Cysteine HCl x H2O | 0.5 | g | ||
|
|
Na2CO3 | 1.5 | g | ||
|
|
Na2S x 9 H2O | 0.2 | g | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Dissolve ingredients except cysteine, carbonate and sulfide. Sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve cysteine, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture and sulfide from a sterile anoxic stock solution prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 6.8 - 7.0, if necessary. | |||||
| 2 Note: After growth has started add sterile 80% H2 and 20% CO2 gas mixture to 0.5 - 1 bar overpressure. | |||||