838 .
GEOANAEROBACTER MEDIUM
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* Medium for Geomobilimonas luticola DSM 24905
Prepare medium without Na-ascorbate.
Growth conditions: anaerobic
30 °C
Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final pH:
6.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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Ferric citrate monohydrate | 10.0 | g | ||
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KH2PO4 | 0.6 | g | ||
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NH4Cl | 0.3 | g | ||
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MgSO4 x 7 H2O | 0.5 | g | ||
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CaCl2 x 2 H2O | 0.1 | g | ||
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Trace element solution SL-11 | 1.0 | ml | ||
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Selenite-tungstate solution | 1.0 | ml | ||
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Na2CO3 | 1.5 | g | ||
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Na-acetate | 0.8 | g | ||
| Na-ascorbate* | 0.8 | g | |||
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Distilled water | 1000.0 | ml | ||
| 1 First dissolve ferric citrate by heating the water under continuous stirring. After cooling to room temperature adjust the pH to 6.0, then add and dissolve the remaining ingredients, except carbonate, acetate and ascorbate. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add acetate and ascorbate from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to inoculation check the medium pH and adjust to 6.8, if necessary. | |||||
Trace element solution SL-11 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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Na2-EDTA x 2 H2O | 5.2 | g | ||
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FeCl2 x 4 H2O | 1.5 | g | ||
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ZnCl2 | 70.0 | mg | ||
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MnCl2 x 4 H2O | 100.0 | mg | ||
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H3BO3 | 6.0 | mg | ||
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CoCl2 x 6 H2O | 190.0 | mg | ||
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CuCl2 x 2 H2O | 2.0 | mg | ||
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NiCl2 x 6 H2O | 24.0 | mg | ||
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Na2MoO4 x 2 H2O | 36.0 | mg | ||
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Distilled water | 1000.0 | ml | ||
| 1 Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 ml. | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.5 | g |
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Na2SeO3 x 5 H2O | 3.0 | mg |
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Na2WO4 x 2 H2O | 4.0 | mg |
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Distilled water | 1000.0 | ml |