Equipment needed: Autoclave
Hungate tubes
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Casitone | 30.0 | g | 29.703 | - | |
| Yeast extract | 5.0 | g | 4.951 | - | |
| K2HPO4 | 5.0 | g | 4.951 | 28.422 | |
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | 5.0e-4 | 0.002 | |
| L-Cysteine HCl x H2O | 0.5 | g | 0.495 | 2.819 | |
| Sugar mix | 10.0 | ml | - | - | |
| Na2CO3 | 1.5 | g | 1.485 | 14.012 | |
| Meat filtrate | 1000.0 | ml | - | - | |
| 1 Dissolve ingredients, except cysteine, sugar mix, and carbonate, boil medium for 1 min, then cool to room temperature under 80% N2 and 20% CO2 gas atmosphere, add 0.5 g/l L-cysteine-HCl x H2O and dispense under same gas atmosphere into Hungate-type tubes (for strains demanding meat particles put those first into the tube, use 1 part meat particles to 4 or 5 parts fluid). Autoclave at 121°C for 20 min. After autoclaving add the sugar mix from a sterile anoxic stock solution prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust the pH of the medium to pH 7.0, if necessary. | |||||
Media with this solution
| 110a | CMC MEDIUM (N2/CO2) |