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Forensic medicine compound detection kit based on 56 Y chromosome SNP genetic markers

A genetic marker and Y chromosome technology, applied in the field of forensic detection, can solve the problems that the accuracy is affected by the depth and width of sequencing, the sequencing steps are cumbersome, and the cost is high, and it can achieve wide application and promotion value, high resolution, and ensure accurate typing. sexual effect

Active Publication Date: 2021-01-15
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although massively parallel sequencing can find more SNPs, its cost is high, there are sampling errors, and its accuracy is affected by the depth and width of sequencing
However, the steps of next-generation sequencing are cumbersome, costly, time-consuming, and highly dependent on the quality and content of the DNA of the sample to be tested, so it is difficult to be widely used

Method used

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  • Forensic medicine compound detection kit based on 56 Y chromosome SNP genetic markers
  • Forensic medicine compound detection kit based on 56 Y chromosome SNP genetic markers
  • Forensic medicine compound detection kit based on 56 Y chromosome SNP genetic markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0083] The Y chromosome SNP composite detection kit for detection may include the following reagents packaged separately:

[0084] a) Multiplex amplification primer mix. It was obtained by mixing the amplification primers shown in Table 3. The amplification primers were synthesized by Invitrogen Company. The synthetic amplification primers were prepared with ultrapure water to 100pM / μL, and then mixed according to the ratio in Table 7 to make a composite amplification primer. Primer mix.

[0085] b) Compound amplification reaction mixture. The PCR reaction mixture MultiplexPCRMix of Qiagen Company was used in this embodiment.

[0086] c) Multiple single base extension reaction primer mix. It was obtained by mixing the single-base extension reaction primers shown in Table 4, and the single-base extension reaction primers were all synthesized by Invitrogen Company. The synthesized 56 extension primers (co...

Embodiment 2

[0096] Example 2 Using the kit of the present invention to detect 635 irrelevant individual samples.

[0097] 635 unrelated individual samples were tested using the above-mentioned forensic science composite detection kit based on 56 Y chromosome SNP genetic markers. The specific detection process is as follows:

[0098] a. The DNA of 635 samples was extracted by the Chelex-100 method as a composite amplification template;

[0099] b. Using the DNA in step a as a template, 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; 0.3 μL of the composite amplification primer mixture, the composite amplification reaction Mixture 2.5μL, template DNA 0.8μL, add ddH 2 0 to 1.4 μL; 95°C, 15 minutes; 94°C, 30 seconds, 66°C-60°C, 90 seconds, 72°C, 30 seconds, 12 cycles of touchdown amplification (Touchdown-PCR); 94°C, 30 seconds , 60°C, 90 seconds, 72°C,...

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Abstract

The invention belongs to the technical field of forensic medicines, and particularly relates to a compound detection kit in a forensic medicine based on SNP (Single Nucleotide Polymorphism) genetic markers of 56 Y chromosomes. The technical problem to be solved is that identification of ethnic groups C, G, H, J, N and R on Chinese men individual examination materials is carried out by utilizing the SNP genetic markers of the Y chromosomes, and position division is carried out on a Y chromosome evolutionary tree, so that ethnic origin deduction is carried out. According to the technical schemeof the invention, based on the compound detection kit in the forensic medicine based on the SNP genetic markers of the 56 Y chromosomes, the detection kit is prepared from separated and packaged compound amplification primer mixture and multiple single-base elongation reaction primer mixture. By applying the technology of compound amplification and single-base elongation, gene types of the SNP genetic markers of the 56 Y chromosomes of the biological examination materials can be obtained at a time, and samples of the ethnic groups C, G, H, J, N and R in the Chinese men individuals can be accurately attributed to slender branches of the existing commonly-recognized Y chromosome evolutionary tree.

Description

technical field [0001] The invention belongs to the technical field of forensic medicine detection, in particular to a forensic medicine composite detection kit based on 56 Y chromosome SNP genetic markers. Background technique [0002] Single nucleotide polymorphism (Single Nucleotide Polymorphism, SNP) refers to the DNA sequence polymorphism caused by a single base mutation at a specific nucleotide position in the genome, and its form includes the insertion, deletion, conversion, transversion etc. Lander formally proposed in 1996 that SNP is the third generation of new molecular markers following SSR and ISSR. It has the characteristics of low mutation frequency, high genetic stability, wide distribution, rich loci, and easy screening. The above characteristics make SNP quickly occupy the center stage of molecular genetics and become a research hotspot in recent years. [0003] The Y chromosome is a chromosome unique to males. The human Y chromosome is about 5.9kb in l...

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 侯一平谢明坤王正李介男宋凤郎敏罗海玻钱小钦李英碧刘京
Owner SICHUAN UNIV
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