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Group of SNP markers for identifying donkey species, primer group of group of SNP markers and kit prepared by primer group

A primer set and labeling technology, applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve problems such as being easily affected by living environment and activities, spending a lot of time, and face recognition interference, etc. Achieve good generalization, easy operation, and short amplification detection fragments

Inactive Publication Date: 2021-02-05
LIAOCHENG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The facial recognition system used for animals needs to be based on multi-angle facial photos, but the cooperation degree of animals is low, and they are generally lying down or squinting, and the process of facial data entry takes a lot of time
In fact, the individual differences of the same species of animals are even difficult to distinguish with the naked eye, and their appearance is also easily affected by the living environment and activities, which will cause considerable interference to facial recognition

Method used

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  • Group of SNP markers for identifying donkey species, primer group of group of SNP markers and kit prepared by primer group
  • Group of SNP markers for identifying donkey species, primer group of group of SNP markers and kit prepared by primer group
  • Group of SNP markers for identifying donkey species, primer group of group of SNP markers and kit prepared by primer group

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1) Group data acquisition:

[0022] The whole genome resequencing data of 120 donkeys were obtained, and there was no biological relationship between 120 donkey individuals. 60 samples were used to screen SNP sites (training set), and the remaining 60 samples were used as a verification set.

[0023] 2) Reference genome acquisition:

[0024] The latest version of the donkey reference genome file was obtained from the UCSC database (http: / / genome.ucsc.edu / ).

[0025] 3) Raw data format conversion:

[0026] Use fastq-dump to convert the original .sra format to double-ended .fq format, and then use gzip to compress .fa to .fq.gz format.

[0027] 4) Data quality control:

[0028] FastQC (v0.11.5) was used for quality control of the raw data, and the sequencing quality was basically qualified and could be used for subsequent analysis.

[0029] 5) SNP loci typing

[0030] Use GATK to type the sample data to obtain a VCF file, including the location of the SNP site, etc. ...

Embodiment 2

[0049] For each SNP site in Example 1, use the primer design software Primer Premier to design forward primers and reverse primers for PCR reactions in the conserved sequences within 250 bp of the core sequences of the above 10 SNP sites. Primers, as shown in Table 2.

[0050] Table 2. Forward and reverse reference primers for 10 SNP sites

[0051]

[0052] The primer set is designed according to the conserved sequence within 250 bp of both ends of the core sequence of at least one of the SNP sites described in the first aspect of the present invention. The primer set is designed using the conserved sequence within 250bp at both ends of the core sequence of each SNP site to achieve the amplification of each SNP site, thereby ensuring the accuracy of the amplification.

[0053] The primer set is selected from at least one pair of the following: sequences shown in SEQ ID NO:1~SEQ ID NO:20. When performing multiplex PCR reactions, the primer sequences should be chosen with g...

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Abstract

The invention relates to the field of biological information determination, relates to a group of SNP markers for identifying donkey species, a primer group of the group of SNP markers and a kit prepared by the primer group, specially an SNP marker for donkeys and an application, and particularly a primer group for amplifying the SNP marker, a kit, a genotyping method and a genotyping system. A primer pair provided by the invention can realize co-amplification of gene loci at the same time, and can realize multiplex PCR amplification of each gene locus at the same time. The SEQ ID NO: 1 to theSEQ ID NO: 20 contain 10 pairs of primers which respectively correspond to S01-S10. When different detectable markers are connected to different primer groups, the conditions of different SNP loci are reflected by detecting the corresponding markers. The invention relates to a method and equipment used for donkey individual identification and genetic identification.

Description

technical field [0001] The present invention relates to the field of biological information measurement, and relates to a set of SNP markers and its primer set and kit for identifying donkey species, in particular to a SNP marker for donkeys and its application, especially to a method for amplifying the A SNP-marked primer set, a kit, a genotyping method, a genotyping system, methods and equipment that can be used for donkey individual identification and kinship identification. Background technique [0002] For individual identification and paternity identification of donkeys, there are microsatellite marker identification systems, gene locus identification systems, and the like. Among them, Shenyang Agricultural University has constructed a donkey microsatellite 13-fold PCR system for the 13 dinucleotide microsatellite sites recommended by the International Association of Animal Genetics in 2017. The 13 dinucleotide microsatellite loci (AHT4, ASB23, HMS2, HMS3, HMS6, HMS7,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 王长法苗鑫垚祝晓阳高佳阳张瑞涛李玉华刘桂芹刘文强展延栋柴文琼
Owner LIAOCHENG UNIV
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