Method for cloning embryo by nuclear transfer of bovine somatic cells
A technology for cloning embryos and somatic cells, which can be applied to cells modified by introducing foreign genetic material.
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[0043] 1. Sampling: The healthy cattle ovaries collected from the Hongqi Beef Slaughterhouse in Wuhan City, Hubei Province were stored in DPBS at 39°C. The formula is shown in "Contents of the Invention" and brought back to the laboratory within 3 hours;
[0044] 2. Collection and cultivation of bovine oocytes: use a small amount of ovum-absorbing liquid, see the "Contents of the Invention" for the formula, and use a syringe (No. 18 needle) to absorb bovine cumulus cells-oocytes from the 3-8mm follicles on the surface of the bovine ovary Complexes (COCs), the COCs were washed 3 times with DPBS, washed 3 times with mature culture medium (recipe as above), transferred to 100ul mature culture medium micro-droplets and cultured for 22h (cultivation conditions: 39°C, relative 100% humidity, 5% CO 2 incubator);
[0045] 3. Judging the maturity of bovine oocytes: the bovine oocytes that have been cultured in the maturation medium for 22 hours are blown repeatedly for 3 minutes in 0....
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