1243 .
PARADESULFITOBACTERIUM MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
6.8 - 7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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Ferric citrate monohydrate | 13.70 | g | ||
|
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NaCl | 1.00 | g | ||
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MgCl2 x 6 H2O | 0.40 | g | ||
|
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KH2PO4 | 0.20 | g | ||
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NH4Cl | 0.25 | g | ||
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KCl | 0.50 | g | ||
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CaCl2 x 2 H2O | 0.15 | g | ||
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Modified Wolin's mineral solution | 10.00 | ml | ||
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NaHCO3 | 2.50 | g | ||
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Na-pyruvate | 2.50 | g | ||
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Wolin's vitamin solution (10x) | 1.00 | ml | ||
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Na2S x 9 H2O | 0.10 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 First dissolve ferric citrate by heating the water. After cooling to room temperature adjust the pH to 6.0 and dissolve the other ingredients (except bicarbonate, vitamins, pyruvate and sulfide). Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before inoculation add vitamins, pyruvate and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of pyruvate and vitamins should be sterilized by filtration. Adjust pH of complete medium to 6.8 - 7.2, if necessary. | |||||
Modified Wolin's mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
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 |