368 .
MJAIS MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
6.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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NaCl | 30.00 | g | ||
|
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K2HPO4 | 0.90 | g | ||
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KH2PO4 | 0.70 | g | ||
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CaCl2 x 2 H2O | 1.00 | g | ||
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NH4Cl | 1.25 | g | ||
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MgSO4 x 7 H2O | 3.40 | g | ||
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MgCl2 x 6 H2O | 4.18 | g | ||
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KCl | 0.33 | g | ||
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NiCl2 x 6 H2O | 5.00 | mg | ||
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Na2SeO3 x 5 H2O | 0.50 | mg | ||
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Fe2(SO4)3 x n H2O | 0.01 | mg | ||
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H2WO4 | 5.00 | mg | ||
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FeSO4 x 7 H2O | 0.02 | g | ||
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Ferric citrate | 0.02 | g | ||
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Trace mineral solution | 10.00 | ml | ||
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Trace vitamins | 1.00 | ml | ||
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NaHCO3 | 1.00 | g | ||
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Sulfur
(powder) |
10.00 | g | ||
|
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Na2S x 9 H2O | 0.50 | g | ||
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Resazurin | 1.00 | mg | ||
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Distilled water | 1000.00 | ml | ||
| 1 Mix ingredients, except Trace vitamins, NaHCO3, sulfur and Na2S x 9H2O, adjust pH to 5.5, and autoclave. Filter-sterilize Trace vitamins and 5% NaHCO3 solution and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Separately autoclave 5% Na2S x 9H2O solution under a N2 atmosphere. Distribute the medium into culture vessels containing appropriate amount of sulfur (e.g., 20 ml of medium in 120 ml serum bottles containing 0.6 g sulfur) under a stream of H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Aseptically and anaerobically add the sterile Na2S x 9H2O solution. Pressurize the inoculated bottle to 200 kPa H2-CO2 (4:1, v/v). | |||||
Trace mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
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MnSO4 x n H2O | 0.50 | g | ||
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CoSO4 x 7 H2O | 0.50 | g | ||
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ZnSO4 x 7 H2O | 0.18 | g | ||
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CuSO4 x 5 H2O | 0.01 | g | ||
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AlK(SO4)2 | 0.02 | g | ||
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H3BO3 | 0.01 | g | ||
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Na2MoO4 x 2 H2O | 0.01 | g | ||
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SrCl2 x 6 H2O | 0.01 | g | ||
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NaBr | 0.01 | g | ||
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KI | 0.01 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals. Adjust final pH to 7.0. | |||||
Trace vitamins #
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 pantothenate | 5.0 | mg |
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Vitamin B12 | 0.1 | mg |
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p-Aminobenzoic acid | 5.0 | mg |
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Lipoic acid | 5.0 | mg |
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Distilled water | 1000.0 | ml |