979 .
PETROBACTER SUCCINIMANDENS MEDIUM
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Equipment needed:
Final pH:
7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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K2HPO4 | 0.3 | g | ||
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KH2PO4 | 0.3 | g | ||
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MgCl2 x 6 H2O | 0.2 | g | ||
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CaCl2 x 2 H2O | 0.1 | g | ||
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KCl | 0.1 | g | ||
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NaCl | 1.0 | g | ||
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NH4Cl | 1.0 | g | ||
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Yeast extract | 1.0 | g | ||
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Wolin's vitamin solution | 5.0 | ml | ||
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Modified Wolin's mineral solution II | 10.0 | ml | ||
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Distilled water | 1000.0 | ml | ||
| 1 When grown aerobically prepare the medium without the vitamins, adjust the pH to 7.0 with 10M KOH, and sterilise for 45 min at 110°C. On cooling add the vitamins from a filter sterilised stock solution. Both Geococcus nitratireducens (DSM 15495) and Petrobacter succinimandens (DSM 15512) are supplied from aerobically grown cultures. | |||||
| 2 Anaerobic growth may be obtained by gassing the medium with N2:CO2 (80:20), to which 10mM KNO3 has been added, and dispensing the medium under N2:CO2 (80:20). Sterilise by autoclaving at 110°C for 45 min , and to the cold medium add the filter sterilised vitamins. A reducing agent is not required. | |||||
Wolin's vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
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Biotin | 2.0 | mg |
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Folic acid | 2.0 | mg |
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Pyridoxine hydrochloride | 10.0 | mg |
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Thiamine HCl | 5.0 | mg |
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Riboflavin | 5.0 | mg |
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Nicotinic acid | 5.0 | mg |
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Calcium D-(+)-pantothenate | 5.0 | mg |
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Vitamin B12 | 0.1 | mg |
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p-Aminobenzoic acid | 5.0 | mg |
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(DL)-alpha-Lipoic acid | 5.0 | mg |
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Distilled water | 1000.0 | ml |
Modified Wolin's mineral solution II #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Nitrilotriacetic acid | 1.500 | g | ||
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MgSO4 x 7 H2O | 3.000 | g | ||
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MnSO4 x H2O | 0.500 | g | ||
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NaCl | 1.000 | g | ||
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FeSO4 x 7 H2O | 0.100 | g | ||
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CoSO4 x 7 H2O | 0.180 | g | ||
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CaCl2 x 2 H2O | 0.100 | g | ||
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ZnSO4 x 7 H2O | 0.180 | g | ||
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CuSO4 x 5 H2O | 0.010 | g | ||
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AlK(SO4)2 x 12 H2O | 0.020 | g | ||
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H3BO3 | 0.010 | g | ||
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Na2MoO4 x 2 H2O | 0.010 | g | ||
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NiCl2 x 6 H2O | 0.025 | g | ||
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Na2SeO3 x 5 H2O | 0.300 | mg | ||
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Distilled water | 1000.000 | 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. | |||||