Multiplex amplification kit for cattle paternity identification and individual identification and application thereof

A paternity identification and compound amplification technology, which is applied in the field of compound amplification kits for cattle paternity identification and individual identification, can solve problems such as inability to identify cattle gender, and achieve improved overall identification ability, high resolution, and typing. accurate effect

Pending Publication Date: 2020-08-21
AGCU SCIENTECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since these loci are only located on autosomes and the number is not large, it is not ruled out that two individuals with the same allelic type may be encountered, and the sex of the cow cannot be distinguished

Method used

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  • Multiplex amplification kit for cattle paternity identification and individual identification and application thereof
  • Multiplex amplification kit for cattle paternity identification and individual identification and application thereof
  • Multiplex amplification kit for cattle paternity identification and individual identification and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] A detection method of a cattle paternity test and a multiple amplification kit for individual identification, comprising the following steps:

[0090] 1. Screening of genetic loci

[0091] According to the requirements of high polymorphism, non-linkage between markers, and easy genotyping, as well as the actual primer test results and locus arrangement requirements, 20 chromosomal STR loci and 1 Amel sex locus were screened out: UMN0108, BM2113, FJ232028 , SPS115, CSRM60, BM067, TGLA122, INRA023, BM1818, ILSTS006, UMN0929, Amelogenin, ETH225, BM1824, TGLA126, G18833, TGLA227, FJ232023, FJ232022, FJ232026, SPS113.

[0092] 2. Arrangement of gene loci

[0093] According to the above 21 loci, a unique locus arrangement and chemical fluorescent dye labeling method are designed: UMN0108, BM2113, FJ232028, SPS115, CSRM60 are the first group, and the fluorescent dye marker is 6-FAM; BM067, TGLA122 , INRA023, BM1818, ILSTS006, UMN0929 are the second group, the fluorescent dye...

Embodiment 2

[0111] The application of a compound amplification kit for cattle paternity testing and individual identification comprises the following steps:

[0112] (1) Take 5 blood spot samples from different cattle: The samples are provided by a public security bureau.

[0113] (2) Utilize the Chelex method to carry out the cattle genomic DNA extraction to the sample, use the kit of the present invention to carry out typing detection to the extracted sample, and compare all the site typing of 5 samples, as shown in Table 4:

[0114] Table 4

[0115] site name sample 1 sample 2 sample 3 Sample 4 Sample 5 UMN0108 72,74 74,74 74,74 84,84 74,74 BM2113 127,133 131,135 125,127 127,133 131,133 FJ232028 150,162 154,162 150,154 150,162 150,162 SPS115 215,219 207,209 207,209 213,215 215,219 CSRM60 250,266 254,264 264,264 256,264 246,250 BM067 117,119 117,119 117,133 117,117 117,123 TGLA122 153,155 155,167 155...

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Abstract

The invention discloses a multiplex amplification kit for cattle paternity identification and individual identification and an application thereof. The kit comprises 21 pairs of specific primers for amplifying 20 cattle STR sites and one Amel sex site, the sequences of the specific primers are shown as SEQ ID NO: 1-SEQ ID NO: 42 in a sequence table, and 20 cattle STR sites and one Amel sex site can be amplified at the same time. The invention provides the multiplex amplification kit for cattle paternity identification and individual identification and the application thereof. The kit comprisesan autosomal STR, a Y chromosome STR and an Amel sex site at the same time, the kit is the kit with the most detection sites in the same type of products, can simultaneously amplify and detect 20 cattle STR sites and one cattle sex site in a single tube, has the characteristics of strong specificity, high resolution, accurate typing, high sensitivity and the like, and can completely meet the requirements of cattle paternity identification, individual identification, sex discrimination and the like.

Description

technical field [0001] The invention belongs to the technical field of forensic genetics, and in particular relates to a compound amplification kit for paternity identification and individual identification of cattle and its application. Background technique [0002] In my country, cattle are mainly divided into three categories: cattle, buffalo and yak. Among them, buffaloes belong to Artiodactyla, Bovidae, and Buffalo, and yaks belong to Artiodactyla, Bovidae, Bovine, and Yak subgenus. In my country, yellow cattle refer to all domestic cattle other than yaks and buffalo, belonging to Artiodactyla, Bovidae. , Bovine species in the genus. Therefore, buffalo, ox and yak belong to two different genera, while ox and yak belong to different subgenera. There are 28 local breeds of yellow cattle in China. In addition, there are some other local groups or breeds. It is the country with the most breeds of cattle in the world. [0003] Short tandem repeat loci (Short Tandem Repeat,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6879C12N15/11
CPCC12Q1/6888C12Q1/6879C12Q2600/156
Inventor 颜海波王全印蒋云海杨俊胡如芳康颖丁咏滇张步史明皓郑卫国
Owner AGCU SCIENTECH
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