1529 .
GEOGLOBUS AHANGARI MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
6.8 - 7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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Ferric citrate monohydrate | 11.20 | g | ||
|
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NaCl | 19.00 | g | ||
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MgCl2 x 6 H2O | 9.00 | g | ||
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MgSO4 x 7 H2O | 0.15 | g | ||
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CaCl2 x 2 H2O | 0.30 | g | ||
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KCl | 0.10 | g | ||
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KH2PO4 | 0.60 | g | ||
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(NH4)2SO4 | 0.10 | g | ||
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NaBr | 0.05 | g | ||
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SrCl2 | 0.02 | g | ||
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Trace element solution SL-10 | 1.00 | ml | ||
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NaHCO3 | 2.50 | g | ||
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Yeast extract | 0.10 | g | ||
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Na-pyruvate | 1.00 | g | ||
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FeCl2 x 4 H2O | 0.20 | g | ||
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Wolin's vitamin solution (10x) | 1.00 | ml | ||
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L-Cysteine HCl x H2O | 0.10 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 First, dissolve ferric citrate in water by heating, adjust to pH 5.8, then cool to room temperature. Add other compounds (except bicarbonate, yeast extract, pyruvate, vitamins, cysteine, and ferrous chloride) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Then add cysteine and ferrous chloride and adjust pH to 6.8 - 7.0 with bicarbonate. Dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before inoculation add yeast extract, pyruvate, and vitamins, from sterile anoxic stock solutions prepared under 100% N2 gas. Stock solutions of pyruvate and vitamins should be sterilized by filtration. Adjust the pH of the complete medium to 6.8 - 7.0. | |||||
| 2 Note: Use 5 - 10% (v/v) inoculum. The color of the medium supernatant changes from orange-brown to yellow-greenish during growth. | |||||
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 | ||
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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 | 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. | |||||
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 |