Equipment needed: Autoclave
Hungate tubes
Filter
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
|
Ferric citrate
(Aldrich F6129) |
2.40 | g | 2.374 | 9.692 | |
| (NH4)2SO4 | 0.20 | g | 0.198 | 1.497 | |
| KH2PO4 | 3.00 | g | 2.967 | 21.805 | |
| MgSO4 x 7 H2O | 0.50 | g | 0.495 | 2.007 | |
| CaCl2 x 2 H2O | 0.25 | g | 0.247 | 1.682 | |
|
Yeast extract
(OXOID) |
0.50 | g | 0.495 | - | |
| Allen's trace element solution | 10.00 | ml | - | - | |
| FeCl2 x 4 H2O | 0.26 | g | 0.257 | 1.294 | |
| Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
| L-Cysteine HCl x H2O | 0.04 | g | 0.04 | 0.225 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 First dissolve ferric citrate in water by heating and adjust to pH 6.0, then add other medium ingredients except ferrous chloride, vitamins and cysteine. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add ferrous chloride (dissolved in 0.1 N HCl), vitamins (sterilized by filtration) and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to inoculation check pH and adjust to 6.0, if necessary. | |||||
Media with this solution
| 1604 | ATHALASSOTOGA MEDIUM |