1
Solutions A, B and D are sterilized separately by autoclaving at 121°C for 15 min under 100% N2 gas atmosphere. Solution C is sterilized by filtration 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.
2
Note: A small amount of white precipitate forms after autoclaving, but this has no negative effect on growth.
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