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Main sol. 193a

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Internal Solution-ID: S337

Equipment needed: Autoclave Gassing station Hungate tubes Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 942 ml - -
Solution B 30 ml - -
Solution C 20 ml - -
Solution D 1 ml - -
Solution E 10 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 distributed under the same gas atmosphere in 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 D are prepared under 100% N2 gas atmosphere and sterilized by filtration. Solution E is autoclaved separately under 100% N2 gas. 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 6.8 - 7.0.
2 After inoculation, pressurize vessels with 80% H2 and 20% CO2 gas mixture to 1 bar overpressure.
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 at the beginning. For transfers use 5 - 10% inoculum.
193a PHB/PYRUVATE MEDIUM