579 .
GEOBACTER MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
6.7 - 7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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Ferric citrate monohydrate | 13.7 | g | ||
|
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NH4Cl | 1.5 | g | ||
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NaH2PO4 | 0.6 | g | ||
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KCl | 0.1 | g | ||
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Na-acetate | 2.5 | g | ||
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Modified Wolin's mineral solution | 10.0 | ml | ||
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NaHCO3 | 2.5 | g | ||
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Wolin's vitamin solution (10x) | 1.0 | ml | ||
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Distilled water | 1000.0 | ml | ||
| 1 First dissolve ferric citrate in water by heating and adjust to pH 6.0, then add other medium ingredients except bicarbonate and vitamins. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then add and dissolve bicarbonate and adjust pH to 6.8. Thereafter, dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins to the sterile medium from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration. Adjust pH of complete medium to 6.7 - 7.0, if necessary. | |||||
| 2 Note: Use 5 - 10% (v/v) inoculum. | |||||
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 |