Forensic medicine composite detection kit based on 20 multiple insertion/delection genetic markers

An indel, genetic marker technology, applied in the application field of forensic genetics, can solve the problems of increasing detection cost, increasing the difficulty of compound detection and data analysis, and inability to use mixed sample analysis, saving manpower, high recognition power, and reducing costs. Effect

Active Publication Date: 2013-12-25
SICHUAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The compound amplification of a large number of biallelic genetic markers not only increases the detection cost, but also greatly increases the difficulty of compound detectio

Method used

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  • Forensic medicine composite detection kit based on 20 multiple insertion/delection genetic markers
  • Forensic medicine composite detection kit based on 20 multiple insertion/delection genetic markers
  • Forensic medicine composite detection kit based on 20 multiple insertion/delection genetic markers

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Experimental program
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Effect test

Embodiment 1

[0065] The preparation of embodiment one kit of the present invention

[0066] The Multiplex Indel Genetic Marker Multiplex Assay Kit for Detection includes the following reagents packaged separately:

[0067] a) Amplification primer mixture: It is obtained by mixing the amplification primers shown in Table 1 and synthesized by Shanghai Sangong. Prepare 41 amplification primers with 1×TE to 100uM / μL for use, and then follow the instructions in Table 8. Proportional mixing to make a compound amplification primer mixture;

[0068] b) Amplification reaction mixture 2×Taq PCR Master mix (Bs9293) was purchased from Shanghai Sangong;

[0069] c) Allele typing standard mixture: composed of 63 fluorescently labeled DNA fragments as shown in Table 4, four colors (blue, green, black, and red) fluorescence are FAM, HEX, TAMRA, and ROX .

[0070] After the above reagents were packaged according to their respective routine requirements, a multiple insertion-deletion genetic marker compo...

Embodiment 2

[0074] Example 2 Using the kit of the present invention to detect 149 irrelevant individual specimens of the Han nationality

[0075] Using the above-mentioned forensic medicine composite detection kit based on multiple indel genetic markers, the detection of 149 unrelated individual specimens of Han nationality was carried out. The specific detection process is as follows:

[0076] a. Using the Chelex-100 method to extract genomic DNA from blood samples of 149 Han unrelated individuals as a template for multiplex amplification;

[0077] 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.

[0078]

[0079] Thermal cycle reference for amplification: 95°C for 10min; 94°C for 30sec, 60.5°C for 40sec, 72°C for 50sec (29 cycles); 60°C for 45min.

[0080] c. Capillary electrophoresis

[0081] Take 1 μL of t...

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Abstract

The invention belongs to the field of medicolegal genetics, particularly relates to a forensic medicine composite detection kit based on 20 multiple insertion/delection genetic markers, and aims to achieve the purpose of using multiple insertion/delection genetic markers to perform forensic medicine affiliation identification and individual identification on anthropobiology samples. The adopted technical scheme is that the forensic medicine composite detection kit comprises a complex amplification primer mixture and an allelic ladder mixture which are packed separately; the complex amplification primer mixture totally comprises 41 pieces of amplification primers marked by the 20 multiple insertion/delection genetic markers; the allelic ladder mixture is composed of allelic ladders which are marked by the 20 multiple insertion/delection genetic markers and comprise 63 DNA segments marked by fluorescein. The kit provided by the utility model provides a new technological way for parental right identification and individual identification in forensic medicine.

Description

technical field [0001] The invention belongs to the application field of forensic genetics, in particular to a forensic compound detection kit based on 20 multiple insertion-deletion genetic markers. Background technique [0002] Forensic genetics realizes paternity identification and personal identification through the detection of DNA genetic markers on biological samples. Insertion / deletion polymorphism genetic markers (insertion / deletion polymorphism, Indels) are produced by single point mutations in the human genome, that is, a certain number of nucleotide insertions or deletions at specific base positions in the genome, resulting in polymorphisms formed by changes in the length of DNA fragments Sexual genetic markers. It is widely distributed in the human genome, and its distribution density is second only to single nucleotide polymorphism (single nucleotide polymorphism, SNPs). Indels and SNPs are both biallelic polymorphic genetic markers, and also have the charact...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 侯一平黄健颜静罗海玻魏蔚张霁云利兵李英碧
Owner SICHUAN UNIV
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