Equipment needed: Anaerobic workbench
Filter
Hungate tubes
Autoclave
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| KH2PO4 | 0.30 | g | 0.319 | 2.343 | |
| K2HPO4 | 0.30 | g | 0.319 | 1.83 | |
| NH4Cl | 1.00 | g | 1.063 | 19.867 | |
| NaCl | 40.00 | g | 42.508 | 727.378 | |
| CaCl2 x 2 H2O | 0.20 | g | 0.213 | 1.446 | |
| Yeast extract | 2.00 | g | 2.125 | - | |
|
Trypticase peptone
(BD-BBL) |
2.00 | g | 2.125 | - | |
| Glucose | 3.60 | g | 3.826 | 21.235 | |
| Trace element solution | 1.00 | ml | - | - | |
| Resazurin | 1.00 | mg | 0.001 | 0.005 | |
| Distilled water | 940.00 | ml | - | - | |
| 1 To complete the medium, add the following stock solutions [Filter-sterilize NaHCO3 solution and store under a N2-CO2 (4:1, v/v) gas mixture. The other solutions are autoclaved and stored anaerobically under a N2 atmosphere] to the medium (per 4.7 ml of the medium): | |||||
|
MgCl2 x 6 H2O
(15%) |
0.10 | ml | - | - | |
|
NaHCO3
(5%) |
0.20 | ml | - | - | |
|
L-Cysteine HCl x H2O
(5%) |
0.05 | ml | - | - | |
|
Na2S x 9 H2O
(5%) |
0.04 | ml | - | - | |
| 2 Mix ingredients and adjust pH to 7.2. Distribute the medium into culture vessels (e.g., 4.7 ml of the medium in the Hungate tubes) under a N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. | |||||
Media with this solution
| J706 | SPOROSALIBACTERIUM MEDIUM |