818 .
CHRYSIOGENES MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
7.4 - 7.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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NaCl | 1.20 | g | ||
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KCl | 0.30 | g | ||
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NH4Cl | 0.30 | g | ||
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KH2PO4 | 0.20 | g | ||
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Na2SO4 | 0.30 | g | ||
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MgCl2 x 6 H2O | 0.40 | g | ||
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CaCl2 x 2 H2O | 0.15 | g | ||
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Trace element solution SL-11 | 1.00 | ml | ||
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Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
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NaHCO3 | 0.60 | g | ||
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KNO3 | 0.50 | g | ||
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Wolin's vitamin solution (10x) | 1.00 | ml | ||
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Seven vitamins solution | 1.00 | ml | ||
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Na-acetate | 0.50 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except bicarbonate, nitrate, vitamins and acetate. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, adjust pH to 7.0 - 7.4, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add nitrate, vitamins and acetate 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 atmosphere. Vitamins should be sterilized by filtration. The pH of the complete medium should be adjusted to 7.4 - 7.8, if necessary. | |||||
Trace element solution SL-11 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA x 2 H2O | 5.2 | g | ||
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FeCl2 x 4 H2O | 1.5 | g | ||
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ZnCl2 | 70.0 | mg | ||
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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 | 1000.0 | ml | ||
| 1 Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 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 |
Seven vitamins solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Vitamin B12 | 100 | mg |
|
|
p-Aminobenzoic acid | 80 | mg |
|
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D-(+)-biotin | 20 | mg |
|
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Nicotinic acid | 200 | mg |
|
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Calcium pantothenate | 100 | mg |
|
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Pyridoxine hydrochloride | 300 | mg |
|
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Thiamine-HCl x 2 H2O | 200 | mg |
|
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Distilled water | 1000 | ml |