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Medicolegal composite assay kit based on twenty triallelic SNP (single nucleotide polymorphism) genetic markers

A triallelic and genetic marker technology, applied in the field of forensic composite detection kits for allelic SNP genetic markers, can solve the problems of increased detection costs, limited number of triallelic SNPs, inability to analyze mixed samples, etc., and achieves high efficiency. The effect of energy, classification accuracy, widespread promotion and application value

Active Publication Date: 2012-02-01
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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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  • Medicolegal composite assay kit based on twenty triallelic SNP (single nucleotide polymorphism) genetic markers
  • Medicolegal composite assay kit based on twenty triallelic SNP (single nucleotide polymorphism) genetic markers
  • Medicolegal composite assay kit based on twenty triallelic SNP (single nucleotide polymorphism) genetic markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 Preparation of the kit of the invention

[0073] The three-allelic SNP compound detection kit used for detection may include the following reagents packaged separately:

[0074] a) Compound amplification primer mixture. The amplified primers shown in Table 1 were mixed and synthesized by TaKaRa Biotechnology. The 20 pairs of amplified primers synthesized were configured with ultrapure water to 50pM / μL, and then mixed according to the ratio in Table 6 to make a composite amplification. Primer mix.

[0075] b) Composite amplification reaction mixture. In this example, TaKaRa Biotechnology's PCR reaction mixture One shot La PCRTM Mix was used.

[0076] c) Multiple single base extension reaction primer mixture. The single base extension reaction primers shown in Table 2 were mixed and synthesized by TaKaRa Biotechnology. The 20 synthesized single-base extension reaction primers were configured with ultrapure water to 50pM / μL, and the parameters given in Table 7 were u...

Embodiment 2

[0085] Example 2 Use the kit of the present invention to detect 100 unrelated Han individuals

[0086] Using the above-mentioned forensic compound detection kit based on the tri-allelic SNP genetic marker, we carried out the detection of 100 unrelated Han individuals. The specific detection process is carried out as follows:

[0087] a. Use Chelex-100 method to extract genomic DNA from blood samples of 100 unrelated Han nationality individuals as a template for multiple amplification.

[0088] b. Using the DNA template in step a, the sample is subjected to multiple PCR amplification in the following amplification system using the multiple amplification primer mixture and the multiple amplification reaction mixture.

[0089]

[0090] Thermal cycling parameters for amplification

[0091]

[0092] Cycle 35 times from step 2 to step 4

[0093] 5. 72℃ for 10 minutes

[0094] c. Purification of multiple PCR products, the following is the purification system of each sample amplification product...

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Abstract

The invention belongs to the field of forensic medicine, and in particular relates to a medicolegal composite assay kit based on twenty triallelic SNP (single nucleotide polymorphism) genetic markers. The kit consists of a composite amplification primer mixture, a multiple single-base extension reaction primer mixture, an allelic typing reference material mixture, a composite amplification reaction mixed liquor and a single-base extension reaction mixed liquor, which are packed in a separate way. The kit can quickly and exactly obtain the typing of twenty triallelic SNP (single nucleotide polymorphism) genetic markers of a biologic assay material at one time to ensure the individual source of the assay material and provide a new technical means for the personal identification by a composite amplification reaction and a multiple single-base extension reaction in a single pipe and a capillary electrophoresis platform generally used in a medicolegal heredity lab. Therefore, the kit has good application prospect in the field of forensic medicine.

Description

Technical field [0001] The invention belongs to the field of forensic medicine, and in particular relates to a forensic compound detection kit based on 20 tri-allelic SNP genetic markers for forensic individual identification. Background technique [0002] Forensic DNA analysis realizes the purpose of paternity identification and personal identification through the detection of DNA genetic markers on biological samples. Single Nucleotide Polymorphism (SNP) is a DNA sequence polymorphism caused by a single base mutation at a specific nucleotide position in the genome. It is the most common and widely distributed type of DNA polymorphism in the human genome. . As a third-generation DNA genetic marker, SNP has the characteristics of rich content, genetic stability, low mutation rate, and small amplified fragments. It has special application value in forensic paternity identification and personal identification. [0003] However, SNP in the traditional sense refers to a polymorphism ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 侯一平扎拉嘎白乙拉云利兵颜静翟仙敦罗海玻李英碧
Owner SICHUAN UNIV
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