1
Dissolve ingredients of solution A (except cysteine), bring to the boil, then cool to room temperature under 100% H2 gas. Add cysteine and adjust pH to 7.0, if necessary. Dispense medium under 100% H2 gas atmosphere into anoxic Hungate-type tubes or serum vials vials to 30% of their volume and then autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solution C is prepared under 100% N2 gas atmosphere. Complete the medium by adding appropriate amounts of solutions B and C to the sterile solution A. Adjust pH of complete medium to 8.3 - 8.4, if necessary.
2
After inoculation add sterile H2 gas to 1 - 2 bar overpressure.
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