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Start a timer for 10 min as soon as the first sperm sample is mixed with CPA. Finish the remaining steps within 10 min.

• Using a sharpie marker, label three sets of straws that correspond to each target concentration.
• Load each straw by inserting the cotton end of the straw into the filler. Draw up the desired volume into the straw, then remove the open end of the straw from the liquid. Draw up air into the syringe until the sample in the straw reaches the cotton and is absorbed.
• Seal the straws using the ultrasonic sealer and load them on the Cryokit rack. The thermocouple straw should be placed in the middle of the tray. Fasten all straws down with (2) 0.5mL straws on either end.

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When finished with this step, move on to freezing instructions.

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