Method for identifying sex of mammal embryo

An identification method, mammalian technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of unsatisfactory identification results, embryo damage, etc., achieve high identification efficiency and accuracy, low cost, shorten The effect of reaction time

Pending Publication Date: 2021-11-02
TARIM UNIV
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Problems solved by technology

[0005] However, because some embryonic cells need to be cut out as templates during embryo sex identification, it will cause certain damage to the embryo
In

Method used

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  • Method for identifying sex of mammal embryo
  • Method for identifying sex of mammal embryo
  • Method for identifying sex of mammal embryo

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preparation example Construction

[0032] The FISH method mainly includes the preparation of fluorescein probes, the denaturation and hybridization of probes and target DNA, and observed 3 parts. The probe is first combined with reporting molecules. After hybridization, the fluorescent dye is connected by immunocytochemistry, which produces fluorescence. This method can be identified in cell division and intermediate hybridization. Although the method is specific, it is highly efficient, and the error rate is low, but It takes a long time, the reagent is strict, and the identification cost is high, so it is difficult to apply in production practice.

[0033] The PCR method is based on DNA characteristics, adding an appropriate amount of buffer, template DNA, 4 deoxyonucleotide (D NTP) solution, heat-resistant DNA polymerase, Mg2 + and 1 to synthetic primers in a microcencin. Using DNA in vitro 90-95 ° C high temperature solution spiral denaturing double-stranded cleaved chain, at low temperature (60 ° C), the prime...

Embodiment 1

[0051] The technical solution of this embodiment is:

[0052] (1) Get embryos: including living embryos or chemical embryos and frozen embryos, gender identification.

[0053] (2) Acquisition of genomic DNA of embryonic cells.

[0054] (3) Design and synthesis of primers.

[0055] Male special gene primer sequence: upstream 5'-TTGCCCCCTCCACATACAG-3 '; downstream 5'-agccaatgttaccctatcgtg-3'

[0056] Inner-term gene primer sequence: upstream 5'-ggtccacatgcctcccaag-3 ';

[0057] Downstream 5'-tccctttcctcaggggcctt-3 '

[0058] (4) QPCR reaction: In the PCR reaction system, an excess SYBR fluorescent dye is added;

[0059] Each PCR reaction system 10 μL: 2 × S6 Universal Sybr qpcr Mix 5 μL, upper and downstream primers were 0.25 μL, DNA template 1 μL, DD H 2 O 3.5 μL.

[0060] Q-PCR reaction amplification conditions: 95 ° C Prerequisitability 30s, 95 ° C denaturation 10S, 65 ° C annealing extension 20s, 40 cycles.

[0061] (5) Analysis of amplification curve and gender identification. ...

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Abstract

The invention relates to a method for identifying the sex of a mammal embryo. The method for identifying the sex of the mammal embryo comprises the following steps: (1) obtaining the embryo; (2) obtaining genome DNA of the embryonic cells; (3) designing and synthesizing primers; (4) performing QPCR reaction: adding a fluorescent dye into a PCR reaction system; and (5) amplifying the curve and analyzing, and identifying the sex of the embryo. The method for identifying the sex of the mammal embryo, disclosed by the invention, adopts the real-time fluorescent quantitative PCR method, is simple in design, only needs a pair of primers, can analyze and detect the specificity of amplification reaction through a melting point curve, and has the advantages of high sensitivity, short time consumption, low initial cost, good universality, high automation degree and the like.

Description

Technical field [0001] The present invention belongs to the field of biotechnology, and more particularly to a gender identification method of mammalian embryos. Background technique [0002] The study of gender decisions and control of mammals is a major topic of molecular embryology, which has important theoretical significance and practical application value. The mastery and control of livestock gender is to significantly improve the biotechnology of livestock breeding efficiency. Gender control methods, including X, Y sperm separation, embryonic gender identification, and control insemination environments. X, Y Sperm Separation method will damage sperm in the separation process, especially for more fragile sheep sperm, and the effect is not ideal; and the price is more expensive, and the general laboratory is difficult to configure. Adjust the internal environment of the motherstone uterus, that is, the environments and environments need to further study and improve. Embryo G...

Claims

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Application Information

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IPC IPC(8): C12Q1/6879C12Q1/686
CPCC12Q1/6879C12Q1/686C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 高庆华方翟陶维昆
Owner TARIM UNIV
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