197 .
JCM MEDIUM No. 197
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KH2PO4 | 0.75 | g | ||
|
|
K2HPO4 | 0.75 | g | ||
|
|
MgCl2 x 6 H2O | 0.36 | g | ||
|
|
NH4Cl | 1.00 | g | ||
|
|
Yeast extract
(BD-Difco) |
0.10 | g | ||
|
|
Polypeptone
(Nihon Pharm. Co.) |
0.10 | g | ||
|
|
Sodium acetate | 2.50 | g | ||
|
|
Trimethylamine-HCl | 8.00 | g | ||
|
|
Trace mineral solution | 9.00 | ml | ||
|
|
Trace vitamins | 10.00 | ml | ||
|
|
NaHCO3 | 0.80 | g | ||
|
|
Na2S x 9 H2O | 0.50 | g | ||
|
|
L-Cysteine HCl x H2O | 0.50 | g | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 981.00 | ml | ||
| 1 Mix ingredients except NaHCO3, Trace vitamins, Na2S x 9H2O and cysteine x HCl x H2O, and autoclave under a N2-CO2 (80:20) atmosphere. Filter-sterilize NaHCO3 as an 8% solution and Trace vitamins while gassing with N2. Separately autoclave Na2S x 9H2O and cysteine x HCl x H2O as 5% solutions under a N2 atmosphere. Aseptically and anaerobically add NaHCO3 and Trace vitamins to the medium under a N2-CO2 (80:20) atmosphere. Prior to inoculation, add the sterile Na2S x 9H2O and cysteine x HCl x H2O solutions, and adjust pH to 6.8. | |||||
Trace vitamins #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
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 |
Trace mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 4.500 | g | ||
|
|
FeCl2 x 4 H2O | 0.400 | g | ||
|
|
MnCl2 x 4 H2O | 0.100 | g | ||
|
|
CoCl2 x 6 H2O | 0.170 | g | ||
|
|
ZnCl2 | 0.100 | g | ||
|
|
CaCl2 x 2 H2O | 0.020 | g | ||
|
|
H3BO3 | 0.019 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.010 | g | ||
|
|
NiCl2 x 6 H2O | 0.030 | g | ||
|
|
Distilled water | 1000.000 | ml | ||
| 1 Adjust pH to 7.0 with 1 N KOH. | |||||