1304 .
SULFURITALEA MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Final pH:
6.7 - 7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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MgCl2 x 6 H2O | 0.2 | g | ||
|
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CaCl2 x 2 H2O | 0.1 | g | ||
|
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NaNO3 | 1.7 | g | ||
|
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(NH4)2SO4 | 0.1 | g | ||
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KH2PO4 | 0.1 | g | ||
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KCl | 0.1 | g | ||
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Trace element solution SL-10 | 1.0 | ml | ||
|
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Selenite-tungstate solution | 1.0 | ml | ||
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Na2S2O3 x 5 H2O | 0.4 | g | ||
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NaHCO3 | 5.0 | g | ||
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Wolin's vitamin solution | 1.0 | ml | ||
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Distilled water | 1000.0 | ml | ||
| 1 Dissolve ingredients except trace elements, thiosulfate, bicarbonate and vitamins, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to remove dissolved oxygen and to saturate the solution with CO2. Dispense solution under 80% N2 and 20% CO2 gas atmosphere in vials suitable for anaerobic cultures (e.g., Balch-type tubes) to 30% of volume. Close vials with butyl rubber septa and autoclave. Add trace element solution, selenite-tungstate solution and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. The vitamin solution should be sterilized by filtration. Adjust pH of the complete medium to 6.7 - 7.0, if necessary. | |||||
| 2 After inoculation, add 1 bar overpressure of sterile 100% H2 gas. | |||||
| 3 Note: Use at least 10% (v/v) as inoculum. | |||||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%) |
10.0 | ml | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
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ZnCl2 | 70.0 | mg | ||
|
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
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H3BO3 | 6.0 | mg | ||
|
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CoCl2 x 6 H2O | 190.0 | mg | ||
|
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CuCl2 x 2 H2O | 2.0 | mg | ||
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NiCl2 x 6 H2O | 24.0 | mg | ||
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Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
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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-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.5 | g |
|
|
Na2SeO3 x 5 H2O | 3.0 | mg |
|
|
Na2WO4 x 2 H2O | 4.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Wolin's vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
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Pyridoxine hydrochloride | 10.0 | mg |
|
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Thiamine HCl | 5.0 | mg |
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Riboflavin | 5.0 | mg |
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Nicotinic acid | 5.0 | mg |
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Calcium D-(+)-pantothenate | 5.0 | mg |
|
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Vitamin B12 | 0.1 | mg |
|
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p-Aminobenzoic acid | 5.0 | mg |
|
|
(DL)-alpha-Lipoic acid | 5.0 | mg |
|
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Distilled water | 1000.0 | ml |