Top layer
Internal Solution-ID: S4498
Equipment needed: Autoclave
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Modified Wolfe's mineral solution | 100.000 | ml | - | - | |
| Trace minerals (see Medium No. 197 | 1.000 | ml | - | - | |
| MES | 0.195 | g | 1.931 | 9.889 | |
| NaHCO3 | 0.042 | g | 0.416 | 4.95 | |
| Agarose | 0.150 | g | 1.485 | 2.356 | |
| 1 Adjust pH to 6.2. | |||||
| 2 Autoclave both layer solutions. Shortly after autoclaving, pipette the bottom layer into each culture vessel (typically 0.75 ml in 10 ml glass vials, use pipette tips that have been cut to enlarge the opening). The bottom layer is allowed to cool for at least 30 min to ensure its solidification. Cool the top layer to 35 - 40C, add 1 ml Trace vitamins (see Medium No. 197) and adjust pH to 6.1 - 6.4 by bubbling with sterile CO2 gas. Slowly overlay the top layer onto the solidified bottom layer (3.75 ml in the 10 ml vial). Allow to solidify the top layer a minimum of 3 hr to a maximum of overnight. To inoculate, 10 �l of the inoculum is drown into a pipette tip and inject just above the bottom layer. Seal vials with butyl rubber stoppers and replace the gas phase with a N2-CO2-O2 (8:1:1, v/v) gas mixture. | |||||
Media with this solution
| J628 | FRESHWATER IRON-OXIDIZING BACTERIA MEDIUM |