Bama miniature pig somatic cell cloning method
A technology of somatic cell cloning and Bama pig, which is applied in the field of animal embryo biology and animal reproduction, can solve the problems of not finding the same or similar reports, etc.
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[0072] 1. Collection and In Vitro Maturation of Porcine Oocytes
[0073] Common commercial pig ovaries were collected from local slaughterhouses, placed in 0.9% saline at 30-37°C, transported back to the laboratory within 5 hours, and washed with saline for 3-5 times. Use a 10ml syringe with a 12-gauge needle to extract oocytes with a diameter of 2-6ml follicles on the surface of the ovary, and use a glass pipette to absorb oocytes (COCs) with more than 2 layers of cumulus cells and uniform cytoplasm under a solid microscope. Wash 3 times in egg wash and 2 times in maturation solution. Move the oocytes into the mature liquid droplet equilibrated in the incubator for at least 4h, cover with embryo grade mineral oil, culture for 24h, and then transfer to the mature liquid drop without FSH and LH to continue culturing for 18-20h. The incubator conditions are: 5% CO2 air, maximum relative saturated humidity, 39°C. Then cumulus cells were removed with 0.1% hyaluronidase, and the ...
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