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Main sol. 195d

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Internal Solution-ID: S395

Equipment needed: Gassing station Hungate tubes Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 952 ml - -
Solution B 30 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 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. Solution C is autoclaved under 100% N2 gas. Solutions D and E are prepared under 100% N2 gas atmosphere and sterilized by filtration. To complete the medium solutions B to E are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 7.1 - 7.4.
2 After inoculation add 1 bar overpressure of sterile 80% H2 and 20% CO2 gas mixture.
195d KNH MEDIUM