Main sol. 935a
Internal Solution-ID: S1925
Equipment needed: Hungate tubes
Filter
Autoclave
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NH4Cl | 1.00 | g | 0.988 | 18.473 | |
| K2HPO4 | 0.08 | g | 0.079 | 0.454 | |
| MgCl2 x 6 H2O | 4.50 | g | 4.447 | 21.872 | |
| CaCl2 x 2 H2O | 0.38 | g | 0.375 | 2.554 | |
| NaCl | 30.00 | g | 29.644 | 507.26 | |
| KNO3 | 0.50 | g | 0.494 | 4.887 | |
| Trace element solution SL-10 | 1.00 | ml | - | - | |
| Selenite-tungstate solution | 1.00 | ml | - | - | |
| Yeast extract | 2.00 | g | 1.976 | - | |
| Na-pyruvate | 0.50 | g | 0.494 | 4.49 | |
| Na2CO3 | 1.50 | g | 1.482 | 13.985 | |
| Wolin's vitamin solution | 10.00 | ml | - | - | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients (except carbonate, pyruvate and vitamins), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30-45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. | |||||
| 2 Add pyruvate and vitamins from anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. The pyruvate and vitamins solutions are sterilized by filtration. | |||||
| 3 The pH of the complete medium should be 6.5 - 6.7. | |||||