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Solution A is sterilized by autoclaving at 121°C for 15 min under a 100% N2 gas atmosphere. Solutions B and D are sterilized by filtration under a 100% N2 gas atmosphere. Solution C is sterilized by autoclaving under an atmosphere of 80% N2 and 20% CO2 gas mixture. Appropriate amounts of solutions B to D are added to the sterile solution A in the sequence as indicated. Check the pH of the complete medium and adjust it to 7.0 - 7.2, if necessary.
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