1297 .
NATRONIELLA SULFIDIGENA MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Water bath
Final gas composition:
100 % N2
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Final pH:
9.8 - 10.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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NaHCO3 | 25.00 | g | ||
|
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Na2CO3 | 140.00 | g | ||
|
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NaCl | 15.00 | g | ||
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K2HPO4 | 1.00 | g | ||
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NH4Cl | 0.20 | g | ||
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MgCl2 x 6 H2O | 0.20 | g | ||
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Yeast extract | 0.20 | g | ||
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Trace elements solution (Pfennig & Lippert,1966) | 1.00 | ml | ||
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Selenite-tungstate solution | 1.00 | ml | ||
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Glycerol | 2.00 | g | ||
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Wolin's vitamin solution (10x) | 1.00 | ml | ||
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Sulfur
(powdered) |
2.00 | g | ||
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Na2S x 9 H2O | 0.25 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 Dissolve hydrogen carbonate, carbonate, sodium chloride, and hydrogen phosphate, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense solution under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add ammonium chloride, magnesium chloride, yeast extract, trace elements, glycerol, vitamins, and sulfide from a sterile anoxic stock solution prepared under 100% N2 gas. The vitamin solution should be sterilized by filtration. Sulfur is sterilized by steaming for 3 hours on each of 3 successive days (see medium 35) and added aseptically to the sterile medium as powder while retaining anoxic conditions. Adjust the pH of the complete medium to 9.8 - 10.0, if necessary. | |||||
Trace elements solution (Pfennig & Lippert,1966) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
EDTA | 5.00 | g | ||
|
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FeSO4 x 7 H2O | 2.20 | 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 | 0.03 | g | ||
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CoCl2 x 6 H2O | 0.20 | g | ||
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CuCl2 x 2 H2O | 0.03 | g | ||
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NiCl2 x 6 H2O | 0.03 | g | ||
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Na2MoO4 x 2 H2O | 0.03 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 pH 3.0-4.0 | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.5 | g |
|
|
Na2SeO3 x 5 H2O | 3.0 | mg |
|
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Na2WO4 x 2 H2O | 4.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
|
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 |