Main sol. J448
Internal Solution-ID: S4196
Equipment needed: Autoclave
Filter
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
|
Sea Salt
(Sigma) |
30.00 | g | 3000 | - | |
| Mg(NO3)2 x 6 H2O | 2.54 | g | 254 | - | |
| KH2PO4 | 0.30 | g | 30 | 220.449 | |
| NaHCO3 | 1.00 | g | 100 | 1190.38 | |
| MES | 1.95 | g | 195 | 998.771 | |
|
Sulfur
(powder) |
12.00 | g | 1200 | 37418.1 | |
| Trace minerals | 10.00 | ml | - | - | |
| Trace vitamins | 0.50 | ml | - | - | |
| Na2S x 9 H2O | 0.50 | g | 50 | 208.175 | |
| Resazurin | 1.00 | mg | 0.1 | 0.436 | |
| 1 Mix ingredients except sulfur, NaHCO3 and Na2S x 9H2O, adjust pH to 6.5 and autoclave. Filter-sterilize 8% 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. Aseptically distribute the medium into culture vessels containing appropriate amount of sulfur (e.g., 20 ml culture medium in 120 ml serum bottle containing 0.24 g sulfur) under a H2-CO2 (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Prior to inoculation, add the sterile Na2S x 9H2O solution and readjust pH to 6.5, if necessary. Pressurize the inoculated bottles to 200 kPa H2-CO2 (4:1, v/v). | |||||
Media with this solution
| J448 | KA22 MEDIUM |