503b .
FWM MEDIUM (PERCHLORATE)
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* Medium for Acidovorax sp. DSM 14692
Omit solution E and acetate. Add 0.85 g/l NaNO3 to solution A and use 1.00 g/l pivalic acid as substrate.
Growth conditions: 28 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
7.0 - 7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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Solution A | 950 | ml | ||
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Solution B | 30 | ml | ||
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Solution C | 10 | ml | ||
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Solution D | 1 | ml | ||
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Solution E | 10 | ml | ||
| 1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 – 45 min to make it anoxic, distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions D and E are prepared under 100% N2 gas and sterilized by filtration. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Adjust pH of complete medium to 7.0 - 7.2, if necessary. | |||||
Solution A #
950
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
KH2PO4 | 0.20 | g |
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NH4Cl | 0.25 | g |
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NaCl | 1.00 | g |
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MgCl2 x 6 H2O | 0.40 | g |
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KCl | 0.50 | g |
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CaCl2 x 2 H2O | 0.15 | g |
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Na2SO4 | 0.15 | g |
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Modified Wolin's mineral solution | 10.00 | ml |
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NaNO3 | 0.85 | g |
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Distilled water | 940.00 | ml |
Solution C #
10
| Compound | Amount | Unit | |
|---|---|---|---|
| Na-acetate* | 1.36 | g | |
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Pivalic acid | 1.00 | g |
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Distilled water | 10.00 | ml |
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 | |
|---|---|---|---|
|
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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 |