Two-layer percoll density gradient centrifugal separation method of boar sperms
A density gradient centrifugation, density gradient technology, applied in animal cells, vertebrate cells, germ cells, etc., can solve the problems of affecting sperm fertilization ability, difficult to remove sperm, low sperm motility, etc., to improve poor semen liquefaction and improve pregnancy. rate effect
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[0017] Embodiment 1 The pig semen two-layer percoll density gradient centrifugation method provided by the present invention, the steps and conditions are as follows:
[0018] I. Preparation of Percoll: Commercially available Percoll has a volume concentration of 100%. It needs to be mixed with 1.5M sperm washing solution to reach physiological osmotic pressure, and then diluted to the required concentration with sperm washing solution.
[0019] (1) Preparation of percoll solution with a specific gravity of 1.113g / mL and a volume concentration of 90%: Take 45mL of commercially available Percoll, add 5mL of sperm washing solution, mix well, discard 3.75mL, add 3.75mL of water, mix After uniformity, filter with a 0.22 micron filter membrane for later use;
[0020] (2) The preparation of the percoll solution with a specific gravity of 1.066g / mL and a volume concentration of 45%: use an equal volume of percoll with a volume concentration of 90% to mix with the sperm washing liquid...
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