507b .
CARBOXYDOCELLA MANGANICA MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % CO2
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Final pH:
6.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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KCl | 0.33 | g | ||
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MgCl2 x 6 H2O | 0.52 | g | ||
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CaCl2 x 2 H2O | 0.29 | g | ||
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NH4Cl | 0.33 | g | ||
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KH2PO4 | 0.33 | g | ||
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Trace element solution SL-4 | 10.00 | ml | ||
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Na2CO3 | 2.50 | g | ||
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Na-pyruvate | 1.00 | g | ||
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Yeast extract | 0.20 | 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 carbonate, pyruvate, yeast extract, and vitamins, then sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add carbonate and adjust pH to 6.0 - 6.5, then dispense medium under 100% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add pyruvate, yeast extract and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Stock solutions of pyruvate and vitamins are sterilized by filtration. After completing the medium adjust pH to 6.5 with a sterile anoxic stock solution of 1 N NaOH. | |||||
Trace element solution SL-4 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Na2-EDTA | 0.50 | g | ||
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FeSO4 x 7 H2O | 0.20 | g | ||
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ZnSO4 x 7 H2O | 0.10 | g | ||
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MnCl2 x 4 H2O | 0.03 | g | ||
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H3BO3 | 0.30 | g | ||
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CoCl2 x 6 H2O | 0.20 | g | ||
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CuCl2 x 2 H2O | 0.01 | g | ||
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NiCl2 x 6 H2O | 0.02 | g | ||
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Na2MoO4 x 2 H2O | 0.03 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 First dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds. | |||||
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 |