2
Prepare the medium without the Na2S, yeast extract, glucose, Na2CO3, NaHCO3 and vitamins. Heat the medium in a boiling water bath and cool under a stream of nitrogen. Add the Na2CO3, and NaHCO3 to the cooled medium and check the pH. Dispense into appropriate vessels (Hungate tubes or serum bottles) and autoclave. To the sterile medium add Na2S, yeast extract, and glucose from autoclaved stock solutions. The vitamin solution is filter sterilised and added to the sterilised medium.
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