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Forensic medicine composite detection reagent kit based on 20 triallelic SNP genetic markers and detection method

A tri-allelic and genetic marker technology is applied in the field of forensic compound detection kits for allele SNP genetic markers. It can solve the problems of large gaps in genetic markers, limited number of tri-allelic SNPs, and inability to analyze mixed samples. The effect of widespread promotion and application of value

Inactive Publication Date: 2016-04-06
CENT SOUTH UNIV
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Problems solved by technology

[0003] However, the SNP of the two alleles has its limitations. For example, the amount of information is far from that of STR genetic markers with more alleles. At least 50-60 SNP sites are needed to achieve the accumulation of 13-15 commonly used STRs. Personal identification rate and non-parent exclusion rate
The compound amplification of a large number of biallelic SNPs not only increases the detection cost, but also greatly increases the difficulty of compound detection and data analysis.
Another limitation of biallelic SNPs is that they cannot be used in the analysis of mixed samples
Diallelic SNPs cannot indicate the existence of individuals from multiple sources in mixed samples, so the analysis of mixed samples is powerless
[0005] So far, the number of tri-allelic SNPs that can be applied to forensic personal identification is limited, and it is still impossible to build a system with high recognition ability comparable to the existing STR system

Method used

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  • Forensic medicine composite detection reagent kit based on 20 triallelic SNP genetic markers and detection method
  • Forensic medicine composite detection reagent kit based on 20 triallelic SNP genetic markers and detection method
  • Forensic medicine composite detection reagent kit based on 20 triallelic SNP genetic markers and detection method

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Embodiment 1

[0059] Embodiment one: the preparation of kit of the present invention

[0060] The tri-allelic SNP composite detection kit for detection may include the following reagents packaged separately:

[0061] A. Multiplex amplification primer mix. It is obtained by mixing the amplification primers shown in Table 1. The synthesized 20 pairs of amplification primers are dissolved in water and mixed according to the concentrations in Table 6.

[0062] B. Compound amplification reaction mixture. The PCR reaction mixture OneshotLaPCR of TaKaRaBiotechnology company is used in this embodiment TM Mix.

[0063] C. Multiple single base extension reaction primer mixture. It is obtained by mixing the single-base extension reaction primers shown in Table 2. The synthesized 20 single-base extension reaction primers are dissolved in water and mixed according to the concentrations in Table 7.

[0064] D. Single base extension reaction mixture. Use ABI company's SNaPshotreadyreactionmix single...

Embodiment 2

[0074] Embodiment two: using the method of the present invention to identify 100 irrelevant Han individual test materials

[0075] Using the above-mentioned forensic medicine review test mixture based on the three-allelic SNP genetic marker, we conducted the test of 100 unrelated individual samples, and the specific identification process was as follows:

[0076] A. Genomic DNA was extracted from blood samples of 100 unrelated Han individuals, and the genomic DNA was obtained as a template.

[0077] B. Prepare the amplification primer pools in each group of single-tube multiplex amplification systems respectively, dilute 20 pairs of amplification primers to 50pM / μL with ultrapure water, mix them according to the ratio in Table 6, and use them as primer pools, where each The final concentrations of the primers are shown in Table 6.

[0078] Using the DNA template in step A, use the primer pool and the multiplex amplification reaction mixture in step B to perform multiplex PCR ...

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Abstract

The invention belongs to the technical field of forensic medicine, and particularly relates to a forensic medicine composite detection reagent kit based on 20 triallelic SNP genetic markers and a detection method. The reagent kit is composed of a composite amplification primer mixture, a multiplex single-basic-group extension reaction primer mixture, an allelic ladder mixture, composite amplification reaction mixed liquor and single-basic-group extension reaction mixed liquor which are separately packaged. The invention further discloses the method for carrying out detection through the reagent kit. The method includes the steps of extracting sample nucleic acid, preparing a reaction system for amplification and carrying out electrophoretic analysis. By carrying out a composite amplification reaction and a multiplex single-basic-group extension reaction in a single tube and using a general capillary electrophoresis platform, the types of 20 triallelic SNP genetic markers of a biological sample can be rapidly and accurately obtained at a time, the individual source of the sample can be determined, and a new technological means can be provided for forensic personal identification. The reagent kit has good application prospects in the field of forensic medicine.

Description

technical field [0001] The invention belongs to the field of forensic genetics, in particular to a forensic compound detection kit based on 20 tri-allelic SNP genetic markers for forensic individual identification. Background technique [0002] Single nucleotide polymorphism (Single Nucleotide Polymorphism, SNP), as the third generation of genetic markers, has the characteristics of rich content, genetic stability and low mutation rate, and has special application value in forensic science. At present, several forensic kits with biallelic SNPs have been marketed and applied to forensic personal identification or paternity testing. A large number of experiments and practical application results show that forensic kits with biallelic SNPs can solve many practical problems in forensic science. [0003] However, the SNP of the two alleles has its limitations. For example, the amount of information is far from that of STR genetic markers with more alleles. At least 50-60 SNP site...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156C12Q2600/16
Inventor 扎拉嘎白乙拉廖慧丹蔡继峰刘颖刘艳芳郭亚东闫杰丁艳君
Owner CENT SOUTH UNIV
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