1020 .
SULFURICURVUM (MBM) MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Serum bottle
Final pH:
7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaNO3 | 0.2 | g | ||
|
|
KH2PO4 | 0.2 | g | ||
|
|
NH4Cl | 0.2 | g | ||
|
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MgCl2 x 6 H2O | 0.4 | g | ||
|
|
KCl | 0.2 | g | ||
|
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CaCl2 x 2 H2O | 0.1 | g | ||
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Trace element solution SL-4 | 2.0 | ml | ||
|
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Na2S2O3 x 5 H2O | 2.5 | g | ||
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Distilled water | 1000.0 | ml | ||
| 1 Dissolve ingredients, except sodium thiosulfate, then sparge solution with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 7.0, then distribute medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 100 ml bottles) and autoclave. Prior to inoculation, add thiosulfate from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary. | |||||
| 2 After inoculation, pressurize culture vessels to 0.5 bar overpressure with sterile 100% H2 gas. | |||||
Trace element solution SL-4 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA | 0.50 | g | ||
|
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FeSO4 x 7 H2O | 0.20 | g | ||
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ZnSO4 x 7 H2O | 0.10 | g | ||
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MnCl2 x 4 H2O | 0.03 | g | ||
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H3BO3 | 0.30 | g | ||
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CoCl2 x 6 H2O | 0.20 | g | ||
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CuCl2 x 2 H2O | 0.01 | g | ||
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NiCl2 x 6 H2O | 0.02 | g | ||
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Na2MoO4 x 2 H2O | 0.03 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds. | |||||