521 .
DESULFOMONILE MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
6.8 - 7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 1.0 | g | ||
|
|
Na2SO4 | 2.0 | g | ||
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Na2S2O3 x 5 H2O | 1.0 | g | ||
|
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MgSO4 x 7 H2O | 1.0 | g | ||
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CaCl2 x 2 H2O | 0.1 | g | ||
|
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KH2PO4 | 0.5 | 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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Yeast extract | 1.0 | g | ||
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Sodium resazurin
(0.1% w/v) |
0.5 | ml | ||
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NaHCO3 | 1.0 | g | ||
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Vitamin solution | 10.0 | ml | ||
|
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Na-pyruvate | 2.5 | g | ||
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Na2S x 9 H2O | 0.1 | g | ||
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Distilled water | 1000.0 | ml | ||
| 1 Dissolve ingredients (except bicarbonate, vitamins, pyruvate and sulfide), then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Complete the medium by adding vitamins, pyruvate and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of vitamins and pyruvate are sterilized by filtration. Adjust pH of the complete medium to 6.8 - 7.0. | |||||
| 2 Note: Prior to inoculation 10-20 mg/l sodium dithionite (added from a 5% w/v solution freshly prepared under N2 and filter-sterilized) can be added to the medium to stimulate growth at the beginning. | |||||
Vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Pyridoxine hydrochloride | 5 | mg |
|
|
Riboflavin | 5 | mg |
|
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Calcium pantothenate | 5 | mg |
|
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p-Aminobenzoate | 5 | mg |
|
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Thioctic acid | 5 | mg |
|
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Nicotinamide | 50 | mg |
|
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Biotin | 5 | mg |
|
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Folic acid | 5 | mg |
|
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Vitamin B12 | 5 | mg |
|
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Haemin
(dissolved in 0.1 N NaOH) |
5 | mg |
|
|
1,4-Naphthoquinone
(dissolved in 0.1 N NaOH) |
20 | mg |
|
|
Distilled water | 1000 | ml |
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 | ||
|
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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 |
|
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Distilled water | 1000.0 | ml |