Forensic medicine composite examination kit based on 55 Y-chromosome SNP (Single Nucleotide Polymorphism) genetic markers

A Y-chromosome and genetic marker technology, applied in the determination/inspection of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., to achieve good application prospects and promotion value, high system resolution, and accurate typing.

Active Publication Date: 2018-05-22
SICHUAN UNIV
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  • Application Information

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Problems solved by technology

It has been reported in relevant literature that there are certain differences in mitochondrial SNPs between the northern and southern Han in China, but the difference in Y-SNP has not been reported yet, and about 80% of the Chinese Han population is distributed in the O-M175 haplogroup and its subgroups

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  • Forensic medicine composite examination kit based on 55 Y-chromosome SNP (Single Nucleotide Polymorphism) genetic markers
  • Forensic medicine composite examination kit based on 55 Y-chromosome SNP (Single Nucleotide Polymorphism) genetic markers
  • Forensic medicine composite examination kit based on 55 Y-chromosome SNP (Single Nucleotide Polymorphism) genetic markers

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Experimental program
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Embodiment 1

[0074] The preparation of embodiment 1 kit of the present invention

[0075] The composite detection kit for Han male haplogroup subgroup division based on Y chromosome SNP can include the following reagents packaged separately:

[0076] a) Multiplex amplification primer mix. It is obtained by mixing the amplification primers shown in Table 3 and synthesized by Invitrogen Company. The 55 pairs of synthetic amplification primers are prepared with ultrapure water to 100pM / μL, and then mixed according to the ratio in Table 7 to make a composite amplification Primer mix.

[0077] b) Compound amplification reaction mixture. In this embodiment, the PCR reaction mixture MultiplexPCR Mix from Qiagen Company was used.

[0078] c) Multiple single base extension reaction primer mix. It was obtained by mixing the single-base extension reaction primers shown in Table 4, and was synthesized by Invitrogen Company. The synthesized 55 single-base extension reaction primers were configured...

Embodiment 2

[0088] Example 2 Using the kit of the present invention to detect 1028 unrelated Han male individual specimens

[0089] Using the above-mentioned Han male haplogroup subgroup classification composite detection kit based on 55 Y chromosome SNPs, 1028 Chinese Han male unrelated individuals classified into O-M175 haplogroup were detected. The specific detection process is as follows:

[0090] a. Using the Chelex-100 method to extract DNA from the blood samples of 1028 unrelated Chinese Han male individuals as a template for multiplex amplification;

[0091] b. Using the DNA template in step a, use the composite amplification primer mixture and the composite amplification reaction mixture to perform composite PCR amplification on the sample in the following amplification system. System: multiplex amplification primer mixture 0.28 μL, multiplex amplification reaction mixture 2.5 μL, template DNA 1 μL, add ddH 2 0 to 5 μL; 95°C, 15 minutes; 94°C, 30 seconds, 66°C (falling PCR, eve...

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Abstract

The invention belongs to the technical field of forensic medicines, and concretely relates to a forensic medicine composite examination kit for dividing a China south and north Han male group to O-M175 haplogroup subgroups based on 55 Y-chromosome SNP (Single Nucleotide Polymorphism) genetic markers. The invention aims to solve the technical problem so as to utilize the Y-chromosome SNP genetic markers for further subdividing positions of examined materials from the China south and north Han male group divided to the O-M175 haplogroup on a Y-chromosome evolution tree. The invention provides the technical scheme that the forensic medicine composite examination kit based on the 55 Y-chromosome SNP genetic markers comprises a multiplex amplification primer mixture and a multiple single base extension reaction primer mixture packed separately. The kit provided by the invention applies a multiplex amplification technology and a multiple single base extension technology, so that gene types of the 55 Y-chromosome SNP genetic markers of the biological examined materials can be obtained at the same time, and male samples from the China south and north Han male group divided to the O-M175 haplogroup can be correctly classified to the subgroups of the existing acknowledged O-M175 haplogroup.

Description

technical field [0001] The invention belongs to the technical field of forensic medicine, and in particular relates to a forensic medicine composite detection kit for dividing Han male populations in northern and southern China into O-M175 haplogroup subgroups based on 55 Y chromosome SNP genetic markers. Background technique [0002] Single nucleotide polymorphism (Single Nucleotide Polymorphism, SNP) refers to the DNA sequence polymorphism caused by the variation of a single base sequence in a specific part of the genome, which widely exists in the human genome. Most of the SNPs are biallelic markers, which have the characteristics of small amplified fragments, low mutation rate, stable genetics, and easy automatic typing. They are suitable for degraded DNA samples, phenotype prediction, and ethnic origin inference. [0003] The Y chromosome is a chromosome unique to males. Due to the different genetic patterns, the Y chromosome can be divided into pseudo-autosomal region...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 侯一平郎敏叶懿李介男宋凤谢明坤王正刘京罗海玻李英碧
Owner SICHUAN UNIV
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