Forensic next-generation sequencing kit based on 74 Y chromosome SNP genetic markers

A technology of Y chromosome and genetic markers, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., to achieve high system resolution, high stability, and accurate typing

Active Publication Date: 2019-10-11
SICHUAN UNIV
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, with the improvement of resolution, it is necessary to use more Y-SNP sites, including InDel sites, which means that there is a need for a method that can simultaneously detect SNP sites including InDel sites in large quantities , and this requirement is not met by traditional detection methods such as SNaPshot

Method used

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  • Forensic next-generation sequencing kit based on 74 Y chromosome SNP genetic markers
  • Forensic next-generation sequencing kit based on 74 Y chromosome SNP genetic markers
  • Forensic next-generation sequencing kit based on 74 Y chromosome SNP genetic markers

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Embodiment 1

[0082] The preparation of embodiment 1 kit of the present invention

[0083] The Y chromosome SNP next-generation sequencing kit for detection may include the following reagents packaged separately:

[0084] a) Multiplex amplification primer mix. From the mixture of amplification primers shown in Table 3, synthesized by Thermo Fisher and mixed into a primer pool.

[0085] b) Compound amplification reaction mixture, partially digested primer sequence reagents. In this example, Thermo Fisher IonAmpliSeq was used TM The PCR reaction mix provided by Library Kit 2.0.

[0086] c) Adapter ligation reagents. In this embodiment, Ion XPress of Thermo Fisher is used TM For BarcodeAdapter, mix adapters and tags 1:1 to form a tag-adapter mixture, dilute 2 times for later use. It should be noted that since the main function of the tag is to distinguish different samples, it is not necessary to use the tag when the number of single sequencing samples is only 1.

[0087] d) Library p...

Embodiment 2

[0092] Example 2 Using the kit of the present invention to detect 100 unrelated Han male individual specimens

[0093] Using the above-mentioned forensic next-generation sequencing kit based on Y chromosome SNP genetic markers, 100 unrelated Han adult male volunteers were tested. The specific detection process is carried out as follows:

[0094] a. DNA was extracted from blood samples of 100 irrelevant Han male individuals by kit or salting-out method, and used as a composite amplification template (the DNA extracted by the salting-out method came from the existing DNA in the laboratory);

[0095] b. Using the DNA template in step a, use the above compound amplification reaction mixture and compound amplification primer mixture to carry out compound amplification of the DNA template and 2800M standard DNA in a single tube to construct each sample library. The system is 20 μL: amplification primer mixture 10 μL, compound amplification reaction mixture 4 μL, template DNA 1 μL, ...

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Abstract

The invention belongs to the technical field of forensic medicine, and in particular relates to a forensic next-generation sequencing kit based on 74 Y chromosome SNP genetic markers. The technical problem to be solved in the present invention is to use the next-generation sequencing technology to divide the position of the samples from the Chinese population on the Y chromosome evolutionary tree by using the Y chromosome SNP genetic marker. The technical solution of the present invention is a forensic next-generation sequencing kit based on 74 Y-chromosomal SNP genetic markers, which contains a mixture of 72 pairs of primers for multiple amplification, making simultaneous detection of 74 SNPs a reality. The kit of the present invention applies multiple amplification and next-generation sequencing technologies in a single tube, and can obtain the genotyping of 74 Y chromosome SNP genetic markers of multiple biological samples at one time, and correctly assign male samples from Chinese populations to It is now recognized that the Y chromosome is on the branch of the evolutionary tree.

Description

technical field [0001] The invention belongs to the technical field of forensic medicine, and in particular relates to a forensic next-generation sequencing kit based on 74 Y chromosome SNP genetic markers and its application. Background technique [0002] The Y chromosome is a type of chromosome that only exists in male individuals. Its non-recombination regions of Y chromosome (NRY, Y chromosomes not specified below refer to non-recombination regions) have paternal inheritance, non-recombination, and effective population Small and geographically specific and other characteristics. Because it only exists in male individuals, it plays an important role in forensic science, such as in sexual assault cases where men are suspects, as well as in the identification of the victims of mass disasters and the investigation of missing persons. And because of its characteristics such as paternal inheritance and geographical particularity, it has also become one of the important tools ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888
Inventor 侯一平宋凤高天珍王正钱小钦云利兵李英碧
Owner SICHUAN UNIV
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