Primer composition, kit and method for detecting Y-SNP haplogroup based on next-generation sequencing technology and application of primer composition, kit and method

A next-generation sequencing technology, a Y-SNP technology, is applied in biochemical equipment and methods, recombinant DNA technology, and microbial assay/inspection, etc., and can solve problems such as limited technical throughput and inability to detect Y-SNP sites. Achieve the effect of saving time and cost, accurate inference, and detailed division

Pending Publication Date: 2022-01-28
ACADEMY OF FORENSIC SCIENCE
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Problems solved by technology

However, the throughput of the above technologies is limited, the number of Y-SNPs that can be detected is less than 50, and it is impossible to detect up to hundreds of Y-SNP sites at the same time

Method used

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  • Primer composition, kit and method for detecting Y-SNP haplogroup based on next-generation sequencing technology and application of primer composition, kit and method
  • Primer composition, kit and method for detecting Y-SNP haplogroup based on next-generation sequencing technology and application of primer composition, kit and method
  • Primer composition, kit and method for detecting Y-SNP haplogroup based on next-generation sequencing technology and application of primer composition, kit and method

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Embodiment 1

[0036] This embodiment provides a primer composition and method for high-resolution detection of Y-SNP haplogroups based on next-generation sequencing technology. The steps of specific Y-SNP site screening, primer design and detection method are as follows:

[0037] 1. Y-SNP site screening and primer design;

[0038] According to the proportion of the main haplogroups in the Chinese population, combined with the population of each group, different numbers of downstream branch Y-SNP genetic markers were selected from the Y chromosome phylogenetic tree. In order to ensure the complete structure of the tree, many important node trunk haplogroups are added at the same time. Referring to the 2019 ISOGG website (Version: 15.58) (https: / / isogg.org / tree / HaplogroupTr2019.html), a total of 381 Y-SNP sites for the Chinese population were screened out, forming 359 haplogroups. The detailed information such as the chromosomal position of the selected 381 Y-SNP sites in the human reference g...

Embodiment 2

[0075] This example is the forensic verification of the method for high-resolution detection of Y-SNP haplogroups based on next-generation sequencing technology provided in Example 1.

[0076] In accordance with the requirements of the Scientific Working Group for DNA Analysis Methods (SWGDAM), the sequencing quality, sensitivity, repeatability, accuracy, and applicability of case samples of the method constructed in Example 1 were evaluated and forensic parameters were calculated.

[0077] The average sequencing depth of each Y-SNP site calculated based on 100 samples is as follows figure 2 As shown, the library was constructed smoothly and the sequencing quality was good.

[0078] The results of forensic science verification show that the recommended DNA input amount of this method is at least 1.25ng, but it is still possible to obtain meaningful haplogroup results for micro-sample with extremely small DNA concentration in forensic practice. The sensitivity test results are...

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Abstract

The invention relates to the technical field of biotechnology and genetics, and provides a primer composition, a kit and a method for detecting a Y-SNP haplogroup based on a next-generation sequencing technology and application of the primer composition, the kit and the method. According to the Chinese population haplogroup frequency, 381 Y-SNP loci are screened out by referring to the latest version of ISOGG website (Version: 15.58) in 2019, and a method for Chinese population haplogroup detection is designed by utilizing a next-generation sequencing platform and a multiple PCR amplicon capture technology. The method is high in sensitivity, good in specificity, high in applicability to case samples, good in detection capacity to degraded samples and mixture samples, capable of obtaining high-resolution haplogroup results, careful in haplogroup division and accurate in inference. The method can be applied to family investigation and paternal family source inference of unknown sample geographical sources, and a family source inference technology with high sensitivity, high resolution and strong system efficiency is provided for forensic medicine practice.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and genetics, in particular to a primer composition, a kit, a method and an application thereof for high-resolution detection of Y-SNP haplogroups based on next-generation sequencing technology, which are used to amplify 381 targeted Y-SNP loci of mainstream haplogroups in the Chinese population. Background technique [0002] Y-STR test is a very mature forensic DNA analysis technology, and it also has a very wide range of applications and unique application value in assisting criminal case investigations. However, due to the high mutation rate of Y-STR, the haplotype inconsistency in the same family appeared, which brought troubles to the investigation of the family and hindered the inference of the source of the sample. Therefore, solving the potential confusion caused by Y-STR mutations and correctly judging the geographical, ethnic and even family origins of unknown samples from crime ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6869C12N15/11
CPCC12Q1/6888C12Q1/6869C12Q2600/156C12Q2535/122
Inventor 张素华李成涛李敏陶瑞旸陈安琪刘希玲林源阙庭志赵珍敏朱如心
Owner ACADEMY OF FORENSIC SCIENCE
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