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Forensic next-generation sequencing kit based on 165 y-snps

A Y-SNP, next-generation sequencing technology, applied in the field of forensic genetics, can solve the problems of continuous detection of SNP loci, inability to carry out multiple samples, etc., achieving good application prospects and promotion value, high integrity, and high system resolution. Effect

Active Publication Date: 2021-01-26
SICHUAN UNIV
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Problems solved by technology

[0004] The integrity of the Chinese-specific Y-chromosome phylogenetic tree depends on the complete inclusion of representative SNP sites (including InDel sites) at all levels of branching, but traditional forensic detection methods such as SNaPshot and pyrosequencing cannot be used. Continuous detection of multiple samples and a large number of SNP sites

Method used

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  • Forensic next-generation sequencing kit based on 165 y-snps
  • Forensic next-generation sequencing kit based on 165 y-snps
  • Forensic next-generation sequencing kit based on 165 y-snps

Examples

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Effect test

Embodiment 1

[0093] The preparation of embodiment 1 kit of the present invention

[0094] The Y-SNP composite detection kit for detection can include the following reagents packaged separately:

[0095] 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.

[0096] b) Compound amplification reaction mixture, partially digested primer sequence reagents. In this example, the PCR reaction mixture provided by Thermo Fisher's IonAmpliSeq Library Kit 2.0 was used.

[0097] c) Adapter ligation reagents. In this example, Thermo Fisher’s Ion Xpress BarcodeAdapter was used, and adapters and tags were mixed at a ratio of 1:1 to form a tag-adapter mixture, which was diluted twice 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.

[0098] d) Libr...

Embodiment 2

[0103] Example 2 Using the kit of the present invention to detect 51 unrelated male individual specimens

[0104] Using the above-mentioned forensic next-generation sequencing kit based on 165 Y-SNPs, 51 unrelated Chinese male individual samples were tested. The specific detection process is carried out as follows:

[0105] a. Use a kit or salting-out method to extract DNA from the blood samples of unrelated Chinese males as a template for multiple amplification (the DNA extracted by the salting-out method comes from the existing DNA in the laboratory);

[0106] b. Using the DNA template in step a, use the above composite amplification reaction mixture and composite amplification primer mixture to perform composite amplification of the DNA template and 007 standard DNA in a single tube to construct each sample library. The system is 20 μL: amplification primer mixture 10 μL, multiplex amplification reaction mixture 4 μL, template DNA 1 μL, add nuclease-free water to make up 2...

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Abstract

The invention belongs to the field of forensic genetics, in particular to a forensic next-generation sequencing kit based on 165 Y-SNPs. The technical problem to be solved in the present invention is to use the next-generation sequencing technology to use Y-SNP genetic markers to divide the positions of Chinese male biological specimens of unknown origin on the Y chromosome evolutionary tree with high resolution, so as to infer the origin of their male ancestry . The technical solution of the present invention is a forensic next-generation sequencing kit based on 165 Y-SNP genetic markers, including a mixture of 165 pairs of primers for multiple amplification, so that simultaneous detection of 165 Y-SNPs becomes a reality. The kit of the present invention applies compound amplification and next-generation sequencing technology in a single tube, and can obtain the genotyping of 165 Y-SNP genetic markers of multiple biological samples at one time, and infer the geographical location of male samples from Chinese populations. , ethnic or even family origin.

Description

technical field [0001] The invention belongs to the field of forensic genetics, and specifically relates to a second-generation sequencing reagent for forensic medicine based on 165 Y-SNPs for high-resolution division of the position of Chinese male biological specimens from unknown sources on the Y chromosome evolution tree box. Background technique [0002] The Y chromosome is a type of chromosome that only exists in male individuals, and 95% of its regions are non-recombination regions of Y chromosome (NRY, Y chromosomes not specified below refer to non-recombination regions) with paternal inheritance, The characteristics of non-recombination, small effective group and particularity in biogeography make it play an important role in forensic science, such as sexual assault cases where men are suspects, identification of victims of mass disasters, and investigation of missing persons. play an important role. Y-chromosomal short tandem repeat (Y-STR) has been used to find ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/156
Inventor 侯一平钱小钦王正王梦鸽宋凤郎敏诸虹李介男谢明坤
Owner SICHUAN UNIV
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