Bovine STR composite amplification detection kit, primer composition and application thereof

A primer composition and a technique for amplifying primers, applied in the field of biology

Pending Publication Date: 2022-05-31
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is how to accurately and conveniently solve the problem of cattle individual identification, and / or, the problem of cattle kinship identification

Method used

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  • Bovine STR composite amplification detection kit, primer composition and application thereof
  • Bovine STR composite amplification detection kit, primer composition and application thereof
  • Bovine STR composite amplification detection kit, primer composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] Embodiment 1, the screening of bovine fluorescent STR composite amplification detection kit STR site and the design of primers

[0111] This embodiment provides a bovine fluorescent STR composite amplification detection kit, which includes a five-color fluorescent composite amplification primer system, through the research and optimization of the sequence composition characteristics of the conserved regions at both ends of the core repeat sequence of 16 bovine STR sites, Finally, 14 bovine STR loci (STR loci) were selected as target loci for primer design. The corresponding information of each STR locus (STR locus) is shown in Table 2:

[0112] Table 2 Information of bovine STR loci

[0113] site name Repeat sequence characteristics repeat sequence BM1818 simple (TG)n BM1824 simple (GT)n BM2113 simple (CA)n CSRM60 simple (AC)n CSSM66 simple (AC)n ETH3 compound (GT)nAC(GT) 6

ETH10 simple (AC...

Embodiment 2

[0144] Embodiment 2, multiplex amplification system primer concentration optimization

[0145] The optimization and establishment of the fluorescent STR multiplex amplification primer system is the core content of the development of the bovine fluorescent multiplex amplification kit. After completing the screening and design of single primers, it is necessary to mix each single primer and carry out multiple amplification debugging. Like single-site amplification, it is necessary to exclude primers that produce non-specificity and low amplification efficiency; finally, after initially determining After compound amplification primers for each locus, adjust the primer concentration. In the initial compound amplification, equimolar primers should be added to the reaction system, and then the concentration ratio of each pair of primers should be adjusted according to the reaction results; finally, the sequence and concentration of the primers should be determined through a large num...

Embodiment 3

[0149] Embodiment 3, bovine STR complex amplification buffer reagent system construction

[0150] The complex amplification reagent system determines the sensitivity, stability, amplification ability, amplification efficiency and many other key performance parameters of the reagents. First, based on the primer system prepared in Example 2, PCR buffer optimization and research were carried out. After a large number of PCR component and concentration optimization experiments, the influence of different components on the amplification performance of the system was explored, and a system optimization and evaluation method was established. On the basis of determining the components of each group, it is necessary to carry out the joint application test of each component, optimize the supporting cycle parameters, and finally establish the complex amplification buffer reagent system. The final reaction system is shown in Table 4, and the PCR cycle parameters are shown in Table 5. Show...

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Abstract

The invention discloses a bovine STR composite amplification detection kit, a primer composition and application thereof. In particular to a primer composition capable of simultaneously amplifying 14 STR (short tandem repeat) loci of cattle, the 14 STR loci are respectively TGLA227, BM2113, ETH10, SPS115, ILSTS006, TGLA126, TGLA122, INRA023, BM1818, ETH225, BM1824, CSRM60, HAUT27 and CSSM66, and the primer composition consists of primers with nucleotide sequences respectively as shown in SEQ ID No.1-28. The primer composition for cattle individual identification and paternity identification, the kit and the detection method have the advantages of high sensitivity, good stability, realization of direct sample amplification, and simple use method, and meet the requirements of cattle DNA related case detection and DNA database construction.

Description

technical field [0001] The invention belongs to the field of biotechnology, relates to the technical field of forensic science DNA testing, in particular to bovine STR compound amplification detection kits, primer compositions and applications thereof. Background technique [0002] Commonly known as "cattle" is a family of animals under Artiodactyla, Bovidae, and Bovine subfamily. There are 5 genera in total, including African buffalo and American bison, ox, buffalo and yak. Among them, yellow cattle are a common breed of cattle inherent in China. The number of cattle raised in China ranks first among large livestock or cattle. The breeding areas are almost all over the country. They are mainly used in agricultural areas and semi-agricultural and semi-pastoral areas. It is used for both milk and meat in pastoral areas. It is a very important livestock for farmers and herdsmen in our country. [0003] Short tandem repeats (STR), also known as microsatellite DNA (Microsatelli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2531/113C12Q2525/151C12Q2565/125C12Q2563/107Y02A40/70
Inventor 金鑫张颖
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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