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Main sol. 149

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Internal Solution-ID: S259

Equipment needed: Autoclave Filter Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 970 ml - -
Solution B 20 ml - -
Solution C 1 ml - -
Solution D 10 ml - -
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solution C is prepared under 100% N2 gas atmosphere and sterilized by filtration. Solution D is autoclaved separately under 100% N2 gas. To complete the medium appropriate amounts of solutions B to D are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 7.0 - 7.2.
2 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth of some strains at the beginning. For transfers use 5 - 10% inoculum.
149 MEGADESULFOVIBRIO MEDIUM