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Main sol. 194a

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Internal Solution-ID: S359

Equipment needed: Gassing station Serum bottle Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 932 ml - -
Solution B 1 ml - -
Solution C 30 ml - -
Solution D 20 ml - -
Solution E 1 ml - -
Solution F 10 ml - -
Solution G 10 ml - -
1 Solution A is sparged with 80% N2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min ), then distributed under the same gas atmosphere in anoxic serum vials (e.g., 50 ml medium in 100 ml serum bottles) and autoclaved. Solutions B, D and G are autoclaved separately under 100% N2 gas. Solution C is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solutions E and F are prepared under 100% N2 gas atmosphere and sterilized by filtration. Solutions B to G are added to the sterile, cooled solution A in appropriate amounts in the sequence as indicated. Final pH of the medium should be 7.2 - 7.4.
2 Note: For transfers use 5 - 10% (v/v) inoculum. Incubate tubes in a slanted position.
194a DESULFOTOMACULUM OX39 MEDIUM (XYLENE)