206 .
JCM MEDIUM No. 206
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
6.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Salt base solution | 1000.0 | ml | ||
|
|
Trace minerals | 10.0 | ml | ||
|
|
Citric acid | 5.0 | mg | ||
|
|
Tryptone
(BD-Difco) |
6.0 | g | ||
|
|
Sulfur
(powder) |
6.0 | g | ||
|
|
Na2S x 9 H2O | 0.5 | g | ||
|
|
Resazurin | 1.0 | mg | ||
| 1 Mix ingredients, except sulfur and Na2S x 9H2O, and adjust pH to 6.5 with H2SO4. Sterilize by filtration while gassing with H2-CO2 (80:20) gas mixture. Steam sulfur for 3 hr on each of 3 successive days. Neutralize Na2S x 9H2O as a 5% solution, and autoclave under a N2 atmosphere. Aseptically and anaerobically add sterile sulfur to the medium under a H2-CO2 (80:20) atmosphere. Prior to inoculation, add the sterile Na2S x 9H2O solution. Pressurize inoculated bottles to 100 kPa H2-CO2 (80:20). | |||||
Salt base solution #
| 1 Use Salt base solution of Medium No. 151 with 17.0 g/L (final) NaCl and 2.5 mg/L (final) KI. |
Trace minerals #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
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MnSO4 x n H2O | 0.50 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
CoSO4 x 7 H2O | 0.10 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
ZnSO4 x 7 H2O | 0.10 | g | ||
|
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
|
AlK(SO4)2 | 0.01 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
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Distilled water | 1000.00 | ml | ||
| 1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0. | |||||