Application of SRY (sex determining region of the Y) antibody
An antibody and mammalian technology, applied in the field of cell engineering, can solve the problems of low accuracy, waste of embryos, time-consuming, etc., and achieve the effects of less damage to sperm, avoiding waste, and simple methods
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Embodiment 1
[0036] Example 1: Obtaining sex-controlled embryos through bovine in vitro fertilization
[0037] 1. Collection and pretreatment of ovaries
[0038] Cut off the connective tissue from the collected ovaries, place them in saline at 37°C and bring them back to the laboratory within 2 hours. Wash the ovaries several times with preheated saline and set aside.
[0039] 2. Oocyte Maturation in Vitro
[0040] Select follicles with a diameter of 2-8mm to extract follicular fluid with a disposable syringe. Collect oocytes with intact zona pellucida under a dissecting microscope, wash the collected oocytes three times with egg washing solution, wash once in maturation solution, and then place them in maturation solution at 38-39°C, 5% CO2 in the air and saturated humidity conditions for maturation culture, cultured for 22-28 hours to obtain mature oocytes. The egg washing solution is formulated as TCM-199 9.5g / L, NaHCO 3 2.2g / L, sodium pyruvate 25mg / L, Hepes 4.8g / L, penicillin 100m...
Embodiment 2
[0046] Example 2: Embryo gender identification
[0047] When the number of embryonic cleavages was more than 4, the fertilized embryos incubated with different concentrations of SRY antibodies were used for gender identification by PCR.
[0048] Two pairs of primers were designed according to the SRY gene and the autosomal gene, and the primers are shown in Table 1.
[0049] Table 1 Primer sequences for embryo sex identification
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[0051]
[0052]Embryos were treated before PCR: 3.5 μL of proteinase K was added to the embryos, denatured at 55°C for 30 minutes to lyse the cells, and proteinase K was inactivated at 98°C for 10 minutes. Then use random primers to amplify the template reaction system and the reaction procedure is as follows:
[0053] The PCR amplification template reaction system is:
[0054]
[0055] The PCR amplification template reaction procedure is:
[0056]
[0057] Embryo gender identification was carried out by multiplex PCR, among ...
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