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26 mitochondria SNP genetic markers-based forensic medical composite detection kit

A genetic marker and composite detection technology, applied in the field of forensic science, can solve the problems of limited value and difficulty in wide application of control area sequencing, and achieve accurate typing, wide promotion and application value, and improve the effect of resolution

Active Publication Date: 2014-12-17
SICHUAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, these solutions can only be used as a supplement to the sequencing of the control region, and the value of single application is limited. They cannot solve the problem that the sequencing of the control region is difficult to be widely used

Method used

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  • 26 mitochondria SNP genetic markers-based forensic medical composite detection kit
  • 26 mitochondria SNP genetic markers-based forensic medical composite detection kit
  • 26 mitochondria SNP genetic markers-based forensic medical composite detection kit

Examples

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Effect test

Embodiment 1

[0069] The preparation of embodiment 1 kit of the present invention

[0070] The tri-allelic SNP composite detection kit for detection may include the following reagents packaged separately:

[0071] a) Multiplex amplification primer mix. It is obtained by mixing the amplification primers shown in Table 4 and synthesized by Invitrogen Company. The 22 pairs of synthetic amplification primers are prepared with ultrapure water to 10pM / μL, and then mixed according to the ratio in Table 8 to make a composite amplification Primer mix.

[0072] b) Compound amplification reaction mixture. In this embodiment, PCR reaction mixture Multiplex PCRMix from Qiagen Company was used.

[0073] c) Multiple single base extension reaction primer mix. It was obtained by mixing the single-base extension reaction primers shown in Table 5, and was synthesized by Invitrogen Company. The synthesized 26 single-base extension reaction primers were configured with ultrapure water to 10 pM / μL, and the ...

Embodiment 2

[0082] Example 2 Using the kit of the present invention to detect 200 unrelated Han nationality individual specimens

[0083] Using the above-mentioned forensic medicine compound detection kit based on mitochondrial SNP genetic markers, the detection of 200 unrelated Han nationality individual specimens was carried out. The specific detection process is as follows:

[0084] a. Using the Chelex-100 method to extract DNA from blood samples of 200 unrelated Han individuals as templates for multiplex amplification;

[0085] 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; 10.5 μL of the composite amplification primer mixture, the composite amplification reaction mixture Solution 12.5 μL, template DNA 2 μL, add ddH2O to 25 μL; 94°C for 5 minutes, 94°C for 30 seconds, 60°C for 30 seconds, 72°C for 30 seconds,...

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Abstract

The invention belongs to the technical field of forensic medicine, and particularly relates to a 26 mitochondria SNP genetic markers-based forensic medical composite detection kit which is used for carrying out forensic medical personal identification on the unknown human biological samples by utilizing SNP genetic markers. The 26 mitochondria SNP genetic markers-based forensic medical composite detection kit comprises a multiplex amplification primer mixture, a multiple single base extension reactant mixture and an allele parting standard mixture, wherein the mixtures are separately packaged. According to the kit, the technologies of complex amplification in single tubes and multiple single base extension are adopted, so that the gene parting of the 26 mitochondria SNP genetic markers of the biological samples can be obtained at one time and the forensic medical personal identification can be rapidly carried out.

Description

technical field [0001] The invention belongs to the technical field of forensic medicine, in particular to a forensic medicine compound detection kit based on 26 mitochondrial SNP genetic markers. Background technique [0002] Mitochondria is an organelle that exists in the cytoplasm and has its own genetic material—mitochondrial DNA (mitochondrial DNA; mtDNA). Mitochondrial DNA is the only extranuclear genetic material in a closed double-circle structure. Unlike nuclear DNA, mitochondrial DNA molecules are small, only 16,569 base pairs (basePair, bp); the number of copies in a single cell is large, and each can have 10-1000 mtDNA molecules; the mutation rate of mitochondrial DNA is high, at less than 2 There are many polymorphic sites in the mitochondrial genome of 10,000 base pairs; mitochondrial DNA does not participate in recombination, and each polymorphic site in it is passed down independently in the form of haplotype as a whole, showing maternally inherited traits....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156
Inventor 侯一平颜静任峥罗海玻李英碧
Owner SICHUAN UNIV
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