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Main sol. 739

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Internal Solution-ID: S1512

Equipment needed: Gassing station Hungate tubes Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 952 ml - -
Solution B 10 ml - -
Solution C 1 ml - -
Solution D 10 ml - -
Solution E 20 ml - -
Solution F 10 ml - -
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic and reach a pH of around 6, then distribute under the same gas atmosphere into anoxic Hungate-type tubes and autoclave. Solutions B and C are prepared under 100% N2 gas atmosphere and sterilized by filtration. Solutions D and F are autoclaved separately under 100% N2 gas. Solution E is prepared under 80% N2 and 20% CO2 gas gas atmosphere. To complete the medium appropriate amounts of solutions B to F are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be at 7.0. After combining solutions the medium should equilibrate overnight and a white precipitate should be apparent.
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) just before inoculation may stimulate growth at the beginning. For transfers use 5 - 10% (v/v) inoculum.
739 DESULFONEMA ISHIMOTOI MEDIUM