1
Sparge solution A with 100% N2 gas for 30 - 45 min to make it anoxic, distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solutions B and C are autoclaved separately under 100% N2 gas. Adjust pH of complete medium to 7.2 - 7.4, if necessary.
2
Use 1.10 g/l Na-pyruvate as substrate. Na-pyruvate is added to the autoclaved medium from a sterile anoxic stock solution sterilized by filtration.
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