1058 .
THIOHALOPHILUS MEDIUM
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Equipment needed:
Autoclave
Gassing station
Final pH:
7.5 - 7.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 120.00 | g | ||
|
|
K2HPO4 | 1.50 | g | ||
|
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NH4Cl | 0.50 | g | ||
|
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Trace elements solution (Pfennig & Lippert,1966) | 1.00 | ml | ||
|
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CaCl2 x 2 H2O | 0.05 | g | ||
|
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MgSO4 x 7 H2O | 0.50 | g | ||
|
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Na2S2O3 x 5 H2O | 5.00 | g | ||
|
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NaHCO3 | 5.00 | g | ||
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Seven vitamins solution | 1.00 | ml | ||
|
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Distilled water | 1000.00 | ml | ||
| 1 Dissolve sodium chloride, potassium hydrogenphosphate and ammonium chloride, then sparge solution with 80% N2 and 20% CO2 gas mixture for at least 30 - 45 min to remove dissolved oxygen and to saturate the solution with CO2. Dispense solution in vials suitable for anaerobic cultures (e.g. Balch-type tubes) to 50% volume under air atmosphere. Close vials with butyl rubber septa to prevent free exchange of oxygen with the external atmosphere and autoclave. Add trace elements, calcium chloride, magnesium sulfate, thiosulfate, bicarbonate and vitamins from sterile stock solutions and adjust pH of the medium to 7.5 - 7.8 using a sterile stock solution of sodium carbonate (5% w/v). | |||||
| 2 Note: Use at least 10% (v/v) as inoculum. | |||||
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 | |||||
Seven vitamins solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Vitamin B12 | 100 | mg |
|
|
p-Aminobenzoic acid | 80 | mg |
|
|
D-(+)-biotin | 20 | mg |
|
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Nicotinic acid | 200 | mg |
|
|
Calcium pantothenate | 100 | mg |
|
|
Pyridoxine hydrochloride | 300 | mg |
|
|
Thiamine-HCl x 2 H2O | 200 | mg |
|
|
Distilled water | 1000 | ml |