342 .
METHANOBACTERIUM ALCALIPHILUM MEDIUM
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Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final gas composition:
100 % H2
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Final pH:
8.3 - 8.4
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Solution A | 792 | ml | ||
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Solution B | 210 | ml | ||
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Solution C | 10 | ml | ||
| 1 Dissolve ingredients of solution A (except cysteine), bring to the boil, then cool to room temperature under 100% H2 gas. Add cysteine and adjust pH to 7.0, if necessary. Dispense medium under 100% H2 gas atmosphere into anoxic Hungate-type tubes or serum vials vials to 30% of their volume and then autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solution C is prepared under 100% N2 gas atmosphere. Complete the medium by adding appropriate amounts of solutions B and C to the sterile solution A. Adjust pH of complete medium to 8.3 - 8.4, if necessary. | |||||
| 2 After inoculation add sterile H2 gas to 1 - 2 bar overpressure. | |||||
Solution A #
792
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NH4Cl | 1.0 | g |
|
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MgCl2 x 6 H2O | 0.1 | g |
|
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CaCl2 x 2 H2O | 0.1 | g |
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K2HPO4 | 0.4 | g |
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Na2SeO3 x 5 H2O
(0.01% w/v) |
2.0 | ml |
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Modified Wolin's mineral solution | 10.0 | ml |
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Yeast extract
(OXOID) |
2.0 | g |
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Trypticase peptone
(BD BBL) |
2.0 | g |
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Sodium resazurin
(0.1% w/v) |
0.5 | ml |
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L-Cysteine HCl x H2O | 0.5 | g |
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Distilled water | 780.0 | ml |
Modified Wolin's mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Nitrilotriacetic acid | 1.50 | g | ||
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MgSO4 x 7 H2O | 3.00 | g | ||
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MnSO4 x H2O | 0.50 | g | ||
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NaCl | 1.00 | g | ||
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FeSO4 x 7 H2O | 0.10 | g | ||
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CoSO4 x 7 H2O | 0.18 | g | ||
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CaCl2 x 2 H2O | 0.10 | g | ||
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ZnSO4 x 7 H2O | 0.18 | g | ||
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CuSO4 x 5 H2O | 0.01 | g | ||
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AlK(SO4)2 x 12 H2O | 0.02 | g | ||
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H3BO3 | 0.01 | g | ||
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Na2MoO4 x 2 H2O | 0.01 | g | ||
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NiCl2 x 6 H2O | 0.03 | g | ||
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Na2SeO3 x 5 H2O | 0.30 | mg | ||
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Na2WO4 x 2 H2O | 0.40 | mg | ||
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Distilled water | 1000.00 | ml | ||
| 1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. | |||||