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Main sol. 792

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Internal Solution-ID: S1601

Equipment needed: Gassing station Serum bottle Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 989 ml - -
Solution B 10 ml - -
Solution C 1 ml - -
Solution D 10 ml - -
1 Sparge solution A with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic serum vials (e.g., 20 ml medium in 100 ml serum bottles) and autoclave. Solutions B and D are autoclaved separately under 100% N2 gas atmosphere and solution C is prepared under 100% N2 gas and sterilized by filtration. To complete the medium appropriate amounts of solutions B, C and D are added to solution A. Adjust pH of the complete medium to 5.8 using a sterile anoxic stock solution of 5% (w/v) Na2CO3, if necessary.
2 After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
3 Note: Solution A has to be prepared freshly. It cannot be stored for a long period of time.
792 PYROLOBUS FUMARII MEDIUM