Main sol. J1033
Internal Solution-ID: S5086
Equipment needed: Anaerobic workbench
Hungate tubes
Autoclave
| 1 Mix components, except sulfur, bring to a boil and cool down under a CO2 (100%) gas stream. While gassing, add the following components to the medium: | |||||
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NaCl | 18.000 | g | 17.476 | 299.037 | |
| MgCl2 x 6 H2O | 4.330 | g | 4.204 | 20.678 | |
| NH4Cl | 0.330 | g | 0.32 | 5.99 | |
| KCl | 0.330 | g | 0.32 | 4.298 | |
| KH2PO4 | 0.165 | g | 0.16 | 1.177 | |
| CaCl2 x 2 H2O | 0.033 | g | 0.032 | 0.218 | |
| Trace elements solution | 10.000 | ml | - | - | |
|
Sulfur
(powder) |
5.000 | g | 4.854 | 151.368 | |
| Distilled water | 1000.000 | ml | - | - | |
| Trace vitamins | 10.000 | ml | - | - | |
| NaHCO3 | 2.000 | g | 1.942 | 23.114 | |
| 2 Bubble CO2 for 3 - 4 min and check pH to be 6.5 - 6.8. Readjust pH to the range if necessary. Autoclave under a CO2 gas atmosphere. Add the following solution to the medium (separately autoclaved under N2): | |||||
|
KNO3
(1 M) |
10.000 | ml | - | - | |
| 3 Steam sulfur for 3 hr on each of 3 successive days. Aseptically and anaerobically distribute the medium into culture vessels containing appropriate amount of sulfur (e.g., 10 ml of the medium and 0.05 g sulfur in Hungate tubes) under a CO2 stream and seal with butyl rubber stoppers. | |||||
Media with this solution
| J1033 | ST65 MEDIUM |