Main sol. J280
Internal Solution-ID: S3972
This solution is used in 2 media and was first defined in medium J280.
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Equipment needed: Autoclave
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| KCl | 0.330 | g | 0.324 | 4.34 | |
| MgCl2 x 6 H2O | 2.800 | g | 2.745 | 13.503 | |
| MgSO4 x 7 H2O | 3.400 | g | 3.333 | 13.524 | |
| NH4Cl | 0.250 | g | 0.245 | 4.582 | |
| NaCl | 25.000 | g | 24.51 | 419.401 | |
| K2HPO4 | 0.300 | g | 0.294 | 1.689 | |
|
Yeast extract
(BD-Difco) |
3.000 | g | 2.941 | - | |
|
Tryptone
(BD-Difco) |
3.000 | g | 2.941 | 41.378 | |
| FeSO4 x 7 H2O | 0.025 | g | 0.025 | 0.088 | |
| NaBr | 10.000 | mg | 0.01 | 0.095 | |
| Trace minerals | 10.000 | ml | - | - | |
| Trace vitamins | 10.000 | ml | - | - | |
|
Sulfur
(powder) |
10.000 | g | 9.804 | 305.704 | |
| Na2S x 9 H2O | 0.500 | g | 0.49 | 2.041 | |
| Resazurin | 1.000 | mg | 9.8e-4 | 0.004 | |
| Distilled water | 1000.000 | ml | - | - | |
| 1 Mix ingredients except sulfur and Na2S x 9H2O, adjust pH to 7.0-7.2 and autoclave under a N2 atmosphere. Steam sulfur for 3 hr on each of 3 successive days. Neutralize Na2S x 9H2O as a 5% solution and autoclave under a N2 gas atmosphere. Distribute the medium and sulfur into culture vessels under a N2 stream, and seal with butyl rubber stoppers. Prior to inoculation, add the sterile Na2S x 9H2O solution, and readjust pH to 7.0-7.2, if necessary. | |||||
| 2 Comment: | |||||
| 3 Most of Thermococcus strains will grow in this medium supplemented with 5.0 g/L maltodextrin and omitted sulfur. | |||||