Main sol. J1067
Internal Solution-ID: S5132
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
FeS solution | 50.00 | ml | - | - | |
Modified Wolfe's mineral solution | 50.00 | ml | - | - | |
Agar
, Noble
(BD-Difco) |
1.50 | g | 1.333 | 3.964 | |
1 Top layer: | |||||
Modified Wolfe's mineral solution | 1000.00 | ml | - | - | |
Trace minerals | 10.00 | ml | - | - | |
2 After cooling of the top layer solution, add per liter the following solutions (filter-sterilized): | |||||
Trace vitamins | 10.00 | ml | - | - | |
NaHCO3
(8%) |
5.25 | ml | - | - | |
3 Bottom layer: | |||||
4 Autoclave both layer solutions. Shortly after autoclaving, pipette the bottom layer into each culture vessel (typically 5 ml in 60 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. | |||||
5 Add the top layer solution in 5 to 6 times the volume of the bottom layer to the culture vessels containing the bottom layer (e.g., 25 to 30 ml in 60 ml glass vials with 5 ml of the bottom layer). | |||||
6 Then, cap the culture vessels with silicone rubber stoppers, and replace the gas phase with a N2-CO2 (4:1, v/v) gas mixture. | |||||
7 Comment: The medium can be stored in culture vessels for a certain time by capping with butyl rubber stoppers, instead of silicon rubber stoppers and keeping anoxic in the vessels. Silicone rubber stopper is O2 permeable and creates a mircoaerophilic condition in the culture vessels. |
J1067 | FRESHWATER IRON-OXIDIZING BACTERIA MEDIUM-II |