1011 .
SULFURIMONAS MJ MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
6.7
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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NaCl | 30.00 | g | ||
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K2HPO4 | 0.14 | g | ||
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CaCl2 x 2 H2O | 0.14 | g | ||
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MgSO4 x 7 H2O | 3.40 | g | ||
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MgCl2 x 6 H2O | 4.18 | g | ||
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KCl | 0.33 | g | ||
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NH4Cl | 0.25 | g | ||
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Fe(NH4)2(SO4)2 x 6 H2O | 0.01 | g | ||
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Modified Wolin's mineral solution | 10.00 | ml | ||
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NaHCO3 | 1.50 | g | ||
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Na2S2O3 x 5 H2O | 1.50 | g | ||
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Wolin's vitamin solution (10x) | 1.00 | ml | ||
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Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except bicarbonate, thiosulfate and vitamins), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials up to a volume of 20% and autoclave. Add bicarbonate, thiosulfate and vitamins to the autoclaved medium from sterile anoxic stock solutions. Solutions of vitamins and thiosulfate are sterilized by filtration and stored under N2, whereas the solution of bicarbonate is prepared under 80% N2 and 20% CO2 gas mixture and autoclaved. Adjust pH of complete medium to 6.7. | |||||
| 2 After inoculation pressurize vessels to 0.5 bar overpressure with sterile 80% N2 and 20% CO2 gas mixture and add sterile air in an amount that is equivalent to a volume of 20% of the headspace. | |||||
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. | |||||
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
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Folic acid | 20 | mg |
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Pyridoxine hydrochloride | 100 | mg |
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Thiamine HCl | 50 | mg |
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Riboflavin | 50 | mg |
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Nicotinic acid | 50 | mg |
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Calcium D-(+)-pantothenate | 50 | mg |
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Vitamin B12 | 1 | mg |
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p-Aminobenzoic acid | 50 | mg |
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(DL)-alpha-Lipoic acid | 50 | mg |
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Distilled water | 1000 | ml |