Main sol. J1159
Internal Solution-ID: S5269
Equipment needed: Filter
Autoclave
Anaerobic workbench
| 1 Mix components and adjust pH to 7.5. Bring to a boil and cool down under a N2-CO2 (4:1, v/v) gas stream. Distribute the medium into culture vessels (e.g., 20 ml medium in 60 ml serum bottles) under the same gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, aseptically and anaerobically add per liter the following solutions (autoclaved or *filter-sterilized): | |||||
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| KH2PO4 | 0.14 | g | 0.133 | 0.977 | |
| MgCl2 x 6 H2O | 0.20 | g | 0.19 | 0.934 | |
| CaCl2 x 2 H2O | 0.15 | g | 0.142 | 0.969 | |
| NH4Cl | 0.54 | g | 0.513 | 9.587 | |
|
Casamino acids
(BD-Difco) |
1.00 | g | 0.95 | - | |
| Yeast extract | 1.00 | g | 0.95 | - | |
| Trace minerals | 1.00 | ml | - | - | |
| Resazurin | 0.50 | mg | 4.7e-4 | 0.002 | |
| Distilled water | 1000.00 | ml | - | - | |
|
NaHCO3
(*, 8%) |
30.00 | ml | - | - | |
|
Trace vitamins
(*) |
10.00 | ml | - | - | |
|
L-Cysteine HCl x H2O
(5%) |
6.00 | ml | - | - | |
|
Na2S x 9 H2O
(5%) |
6.00 | ml | - | - | |
| 2 Readjust pH to 7.0, if necessary. | |||||
Media with this solution
| J1159 | MOLLICUTES H-RF3 MEDIUM |