1086 .
DESULFATIRHABDIUM MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
7.0 - 7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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Na2HPO4 x 2 H2O | 0.53 | g | ||
|
|
KH2PO4 | 0.41 | g | ||
|
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NH4Cl | 0.30 | g | ||
|
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CaCl2 x 2 H2O | 0.11 | g | ||
|
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MgCl2 x 6 H2O | 0.10 | g | ||
|
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NaCl | 0.30 | g | ||
|
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Na2SO4 | 2.80 | g | ||
|
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Trace element solution SL-10 | 1.00 | ml | ||
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Selenite-tungstate solution | 1.00 | ml | ||
|
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Yeast extract | 0.02 | g | ||
|
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Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
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Na2CO3 | 1.50 | g | ||
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Na-benzoate | 0.43 | g | ||
|
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Na-crotonate solution (1 M) | 20.00 | ml | ||
|
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Wolin's vitamin solution (10x) | 1.00 | ml | ||
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Na2S x 9 H2O | 0.50 | g | ||
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|
Distilled water | 980.00 | ml | ||
| 1 Dissolve ingredients (except carbonate, benzoate, crotonate, vitamins and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add benzoate, crotonate, vitamins and sulfide form sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. The crotonate and vitamin solutions should be sterilized by filtration. Adjust pH of the complete medium to 7.0 - 7.2. | |||||
| 2 After inoculation pressurize the vessels with sterile 80% N2 and 20% CO2 gas mixture to 0.7 bar overpressure. | |||||
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 | ||
|
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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 | ||
|
|
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 |
Na-crotonate solution (1 M) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Crotonic acid
(ALDRICH 113018) |
86 | g | ||
|
|
NaOH
(10 N) |
100 | ml | ||
|
|
Distilled water | 900 | ml | ||
| 1 Dissolve crotonic acid in 800 ml distilled water, add around 100 ml of 10 N NaOH and adjust pH to around 7. Then add water to reach a volume of 1000 ml. Sterilize by filtration under 100% N2 gas atmosphere. | |||||
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
|
Folic acid | 20 | mg |
|
|
Pyridoxine hydrochloride | 100 | mg |
|
|
Thiamine HCl | 50 | mg |
|
|
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 |
|
|
p-Aminobenzoic acid | 50 | mg |
|
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
|
Distilled water | 1000 | ml |