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- Dilute the semen with the warmed centrifugation buffer at least 1:1. In this case, we are diluting 10 ml of semen (far right tube) with 20 ml of centrifugation buffer (left tube).
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- Using a Pasteur pipette, put 0.5 ml cushion solution below the semen and
centrifugation buffer to make a cushion for sperm to be pelleted on top.The cushion should be distinct from the sperm solution for it to work properly.
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