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PCR primers for sex identification of nyctereutes procyonoides ussuriensis matschie and an identification method thereof

A gender and primer pair technology, applied in DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of false negatives, gender identification specificity or difficulty in accuracy, and achieve the effect of reducing costs

Inactive Publication Date: 2015-07-22
JILIN UNIV +1
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Problems solved by technology

Since the SRY gene only exists on the Y chromosome of male animals, and female individuals do not have this gene, false negative results are prone to occur during sex identification, which brings difficulties to the specificity or accuracy of sex identification

Method used

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  • PCR primers for sex identification of nyctereutes procyonoides ussuriensis matschie and an identification method thereof

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specific Embodiment

[0013] 1 According to the GenBank database co-existing in the canine sex chromosome X chromosome AMELX gene sequence (NC_006621.3) and sex chromosome Y chromosome AMELY gene sequence (KC763835.1) Use Primer5.0 software to design a pair of primers for amplifying the Ussuri raccoon dog Amelogenin gene, one primer in the primer pair is the nucleotide shown in sequence 1 in the nucleotide sequence table, and the primer pair in The other primer is the nucleotide shown in sequence 2 in the nucleotide sequence table;

[0014] 2 Genomic DNA extracted from blood of 118 Ussuri raccoon dogs by phenol / chloroform method, OD 260 / OD 280 The quality of DNA with a ratio between 1.6 and 1.8 can meet the experimental requirements, according to OD 260 Calculate the concentration of the DNA to be tested based on the value and the dilution factor, and make a corresponding dilution, and aliquot at 50ng / μL as the template DNA;

[0015] 3 Genomic DNA was used as a template for PCR am...

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Abstract

PCR primers for sex identification of nyctereutes procyonoides ussuriensis matschie are disclosed. The nucleotide sequences of the PCR primers are shown as a sequence 1 and a sequence 2 in a nucleotide sequence table. An identification method for sex identification of nyctereutes procyonoides ussuriensis matschie by using the PCR primers is also provided. The method includes performing PCR amplification by adopting total DNA of nyctereutes procyonoides ussuriensis matschie to be identified as a template and by using the PCR primers, and determining sex of a nyctereutes procyonoides ussuriensis matschie individual by combining the PCR amplification and an agarose gel electrophoresis technique. The PCR primers and the method are used for sex identification of the nyctereutes procyonoides ussuriensis matschie, and are rapid, simple, convenient and accurate in result.

Description

technical field [0001] The invention belongs to the technical field of biological detection and identification, and in particular relates to a PCR primer for sex identification of Ussuri raccoon dog and an identification method thereof. technical background [0002] Raccoon dog is one of the precious fur animals. As for the gender identification of raccoon dogs, there is no other effective means except the method of external observation. Therefore, people's judgment of the gender of individual raccoon dogs is only identified through clinical observation. However, in the wild, if you only see the feces or blood of wild raccoon dogs, it is difficult to judge the male and female. According to the traditional method, the conclusion can only be made after observing the individual raccoon dogs, otherwise it cannot be identified. Therefore, there is an urgent need for an effective and rapid diagnostic method to identify male and female raccoon dogs. [0003] Animal gender identi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6879
Inventor 李玉梅闫守庆姚纪元白春燕赵志辉
Owner JILIN UNIV
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