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Before autoclaving, the pH of the Medium is adjusted with NaOH to 7.5-8.0. After autoclaving, the filter-sterilized vitamin solution (0.1% volume), and each of the separately autoclaved, concentrated solutions are added to the medium at a final concentration described below. Then a mix gas (80% H2, 20% CO2) is purged for 5 min. Finally, the mix gas is compressed into gas phase (> 80% volume of the tube or bottle) at 3 atm.
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