198 .
JCM MEDIUM No. 198
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
7.4
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KH2PO4 | 0.75 | g | ||
|
|
K2HPO4 | 0.75 | g | ||
|
|
MnCl2 x 4 H2O | 0.36 | g | ||
|
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NH4Cl | 1.00 | g | ||
|
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NaHCO3 | 4.00 | g | ||
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Trace mineral solution | 9.00 | ml | ||
|
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Trace vitamins | 10.00 | ml | ||
|
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Na2S x 9 H2O | 0.50 | g | ||
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L-Cysteine HCl x 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 NaHCO3, Trace vitamins, Na2S x 9H2O and L-cysteine x HCl x H2O, and autoclave under a H2-CO2 (80:20) atmosphere. Filter-sterilize NaHCO3 as 8% solution and Trace vitamins while gassing with N2. Separately autoclave Na2S x 9H2O and L-cysteine x HCl x H2O as 5% solutions under a N2 atmosphere. Aseptically and anaerobically add the NaHCO3 solution and Trace vitamins to the medium under a H2-CO2 (80:20) atmosphere. Prior to inoculation, add sterile the Na2S x 9H2O and L-cysteine x HCl solutions, and adjust pH to 7.4. Pressurize inoculated bottles to 200 kPa H2-CO2 (80:20). | |||||
Trace mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 4.500 | g | ||
|
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FeCl2 x 4 H2O | 0.400 | g | ||
|
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MnCl2 x 4 H2O | 0.100 | g | ||
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CoCl2 x 6 H2O | 0.170 | g | ||
|
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ZnCl2 | 0.100 | g | ||
|
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CaCl2 x 2 H2O | 0.020 | g | ||
|
|
H3BO3 | 0.019 | g | ||
|
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Na2MoO4 x 2 H2O | 0.010 | g | ||
|
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NiCl2 x 6 H2O | 0.030 | g | ||
|
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Distilled water | 1000.000 | ml | ||
| 1 Adjust pH to 7.0 with 1 N KOH. | |||||
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 |
|
|
Riboflavin | 5.0 | mg |
|
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Nicotinic acid | 5.0 | mg |
|
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Calcium pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |