692 .
METHANOSALSUM WeN5 MEDIUM
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Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
9.2 - 9.4
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ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 15.0 | g | ||
|
|
NaHCO3 | 10.0 | g | ||
|
|
KH2PO4 | 0.3 | g | ||
|
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NH4Cl | 0.5 | g | ||
|
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Modified Wolin's mineral solution | 10.0 | ml | ||
|
|
NiCl2 x 6 H2O
(0.1% w/v) |
2.0 | ml | ||
|
|
Yeast extract | 0.5 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | ||
|
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Na2CO3 | 4.0 | g | ||
|
|
Methanol
(50% v/v) |
10.0 | ml | ||
|
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Na2S x 9 H2O | 1.0 | g | ||
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|
Distilled water | 1000.0 | ml | ||
| 1 Dissolve ingredients except carbonate, methanol and sulfide, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add and dissolve carbonate and adjust pH to 9.0 - 9.2, then dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Prior to inoculation add methanol (50% v/v solution) and sulfide from anoxic stock solutions autoclaved under 100% N2 gas. Adjust pH of complete medium to 9.2 - 9.4. | |||||
Modified Wolin's mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
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. | |||||