Main sol. 1637
Internal Solution-ID: S3400
Equipment needed: Hungate tubes
Filter
Autoclave
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
|
Trypticase peptone
(BD BACTO) |
16.0 | g | 15.686 | - | |
|
Yeast extract
(OXOID) |
7.0 | g | 6.863 | - | |
| NaCl | 5.0 | g | 4.902 | 83.88 | |
| L-Cysteine HCl x H2O | 0.5 | g | 0.49 | 2.791 | |
| NaHCO3 | 0.5 | g | 0.49 | 5.835 | |
| Haemin solution | 5.0 | ml | - | - | |
| Vitamin K1 solution | 5.0 | ml | - | - | |
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | 4.9e-4 | 0.002 | |
|
Methanol
(15% v/v) |
10.0 | ml | 7.765 | 242.329 | |
| DL-Dithiothreitol | 1.0 | g | 0.98 | 6.354 | |
| Distilled water | 1000.0 | ml | - | - | |
| 1 Dissolve ingredients except for methanol and dithiothreitol. Adjust pH to 7.5 with 1 N NaOH and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Distribute medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. After sterilization add methanol from a sterile anoxic stock solution (15% v/v) autoclaved under 100% N2 gas atmosphere and dithiothreitol from an anoxic stock solution sterilized by filtration. Before use check the pH of the complete medium and adjust it to 7.6 - 7.8, if necessary. | |||||
| 2 After inoculation, add sterile 80% H2 and 20% CO2 gas mixture to 0.5 bar overpressure. After growth becomes visible overpressure of 80% H2 and 20% CO2 gas mixture can be increased to 1 bar. | |||||
| 3 Note: Use 10% (v/v) as inoculum. | |||||
Media with this solution
| 1637 | METHANOMASSILIICOCCUS LUMINYENSIS MEDIUM |