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Forensic medicine II sequence testing kit based on 74 gama 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 the effects of accurate typing, high system resolution, good application prospects and promotion value

Active Publication Date: 2017-02-15
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • 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 medicine II sequence testing kit based on 74 gama chromosome SNP genetic markers
  • Forensic medicine II sequence testing kit based on 74 gama chromosome SNP genetic markers
  • Forensic medicine II sequence testing kit based on 74 gama 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 particularly relates to a forensic medicine II sequence testing kit based on 74 gama chromosome SNP genetic markers. The technical problem to be solved is to classify detection materials of Chinese group source at the position of a gama chromosome evolution tree by using the gama chromosome SNP genetic markers. The technical scheme of the invention is forensic medicine II sequence testing kit based on 74 gama chromosome SNP genetic markers, which includes a mixture of 72 pairs of primers for recombination and amplification, thus the detection of 74 pieces of SNP at the same time becomes true. The kit applies single pipe internal recombination and amplification and II sequence testing technology, and thus the genetic typing of 74 pieces of gama chromosome SNP genetic markers of multiple biological detection materials can be obtained in one time; besides, male samples of Chinese group source are correctly affiliated to the branch of the recognized gama chromosome evolution 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 Applications(China)
IPC IPC(8): C12Q1/68
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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