1315 .
ANAEROBIC THAUERA MEDIUM
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* Medium for Acidovorax sp. DSM 15015
Replace fatty acid by 3,4-dihydroxybenzoate (0.77 g/l)
Growth conditions: 28 °C
Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
7.1 - 7.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KNO3 | 0.85 | g | ||
|
|
NaHCO3 | 2.52 | g | ||
|
|
CaCl2 | 0.01 | g | ||
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MgSO4 x 7 H2O | 0.50 | g | ||
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MnSO4 | 0.01 | g | ||
|
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NH4Cl | 0.30 | g | ||
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NaCl | 0.05 | g | ||
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Selenite and tungstate solution | 2.00 | ml | ||
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Distilled water | 884.00 | ml | ||
|
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Phosphate solution | 100.00 | ml | ||
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Acetate solution 98.0 g/L | 10.00 | ml | ||
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Vitamin solution | 5.00 | ml | ||
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Trace element solution SL-10 | 1.00 | ml | ||
|
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3,4-dihydroxybenzoate | 0.77 | g/l | ||
| 1 Distribute to gas tight vessels, gas with 90% N2 +10% CO2 to achieve anaerobic conditions and autoclave. After cooling, add the phosphate solution, substrate solution, vitamin solution and trace element solution. After mixing, pH should be 7.1 - 7.5. | |||||
Phosphate solution #
100
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2HPO4 x 2 H2O | 1.45 | g | ||
|
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KH2PO4 | 0.25 | g | ||
|
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Distilled water | 100.00 | ml | ||
| 1 Distribute to gas tight vessels under N2, autoclave. | |||||
Acetate solution 98.0 g/L #
100
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
K-acetate | 9.8 | g | ||
|
|
Distilled water | 100.0 | ml | ||
| 1 Filter sterilize and gas with N2. For strains DSM 12141, DSM 12142, DSM 11243 and DSM 12144, acetate may be replaced by Na-capronate (stock solution 2.76 g/100 ml). | |||||
Vitamin solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Vitamin B12 | 50 | mg | ||
|
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Pantothenic acid | 50 | mg | ||
|
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Riboflavin | 50 | mg | ||
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Pyridoxamine hydrochloride | 10 | mg | ||
|
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Biotin | 20 | mg | ||
|
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Folic acid | 20 | mg | ||
|
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Nicotinic acid | 25 | mg | ||
|
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Nicotine amide | 25 | mg | ||
| alpha-lipoic acid* | 50 | mg | |||
|
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p-Aminobenzoic acid | 50 | mg | ||
|
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Thiamine-HCl x 2 H2O | 50 | mg | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Stir for some hours, filter sterilize the solution. | |||||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%) |
10.0 | ml | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
|
ZnCl2 | 70.0 | mg | ||
|
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
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H3BO3 | 6.0 | mg | ||
|
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
|
Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
|
Distilled water | 990.0 | ml | ||
| 1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. | |||||
Selenite and tungstate solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Distilled water | 1000.0 | ml | ||
|
|
NaOH | 0.5 | g | ||
|
|
Na2SeO3 x 5 H2O | 3.0 | mg | ||
|
|
Na2WO4 x 2 H2O | 4.0 | mg | ||
| 1 First dissolve NaOH, subsequently the metal salts. Distribute to gas tight vessels under N2, autoclave. | |||||