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Main sol. 383b

Internal Solution-ID: S762

Equipment needed: Gassing station Hungate tubes Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 962 ml - -
Solution B 20 ml - -
Solution C 10 ml - -
Solution D 1 ml - -
Solution E 13 ml - -
1 Solution A is sparged with 80% H2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min), then dispensed under the same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclaved. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C and E are autoclaved under 100% N2 gas. Solution D is prepared under 100% N2 gas and filter-sterilized. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 7.0 – 7.2.
2 After inoculation pressurize vials to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
3 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. Incubate all strains in the dark.