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Medicolegal rapid detection kit based on 21 SNP (Single Nucleotide Polymorphism) mitochondrion loci

A technology for detecting kits, mitochondria, applied in the field of forensic science

Inactive Publication Date: 2016-05-04
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this detection method reduces the number of SNP sites and primers, in actual operation, it still takes 6 to 8 hours to complete the entire process of the above detection method

Method used

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  • Medicolegal rapid detection kit based on 21 SNP (Single Nucleotide Polymorphism) mitochondrion loci
  • Medicolegal rapid detection kit based on 21 SNP (Single Nucleotide Polymorphism) mitochondrion loci
  • Medicolegal rapid detection kit based on 21 SNP (Single Nucleotide Polymorphism) mitochondrion loci

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1 Verification of the accuracy of the kit provided by the invention

[0059] 9947A was used as a DNA template to verify the accuracy of the kit of the present invention.

[0060] 0.1ng of 9947A was used as the composite amplification template for detection, and the detection steps were as follows:

[0061] ⑴. Compound amplification reaction:

[0062] Fast Taq Polymerase for BioDev GoldFaster TM Taq.

[0063] Multiplex amplification reaction system, 20 μL in total:

[0064]

[0065] Multiplex amplification procedure:

[0066]

[0067] ⑵. Purification of composite amplification product: same as Example 1

[0068] ⑶. Single base extension reaction:

[0069] Single base extension reaction system, 5 μL in total:

[0070] Purified composite amplification product 1ng

[0071]Single base extension primer mix 0.5 μL

[0072] SNaPshot reaction mixture 1.2 μL

[0073] Make up to 5 μL with ddH2O

[0074] Single base extension reaction procedure:

[0075...

Embodiment 2

[0078] Example 2 Application of the kit provided by the present invention in individual identification

[0079] The following types of samples were collected from 10 unrelated individuals: blood, semen, saliva, teeth, bones, nails, hair, skin, exfoliated cells, and heart, liver, spleen, lung, stomach, kidney, pancreas, brain, intestine, gallbladder Organs, etc., a total of 190 samples of biological samples, using the kit provided by the present invention and the method of use thereof to detect 21 SNP sites of the above 190 samples.

[0080] The detection steps are as follows:

[0081] ⑴. Mitochondrial DNA extraction:

[0082] Blood, semen spots, saliva and hair samples were extracted by Chelex-100 method; tooth and bone samples were extracted by silica bead method; nail samples were extracted by phenol-chloroform method; skin, exfoliated cells, heart, liver, spleen, lung, stomach, PromegaIQ is used for kidney, pancreas, brain, intestine, and gallbladder samples TM Extractio...

Embodiment 3

[0108] Using 9947A as a DNA template, the sensitivity of the kit provided by the invention was determined.

[0109] Use 0.25pg, 0.5pg, 0.001ng, 0.005ng, 0.01ng, 0.1ng, 1ng, 10ng of 9947A as the composite amplification template for detection, and the detection steps are as follows:

[0110] ⑴. Compound amplification reaction:

[0111] The fast Taq polymerase is GoldFasterTMTaq from BioDev Company.

[0112] Multiplex amplification reaction system, 20 μL in total:

[0113]

[0114] Multiplex amplification procedure:

[0115]

[0116] ⑵. Purification of composite amplification product: same as Example 1

[0117] ⑶. Single base extension reaction:

[0118] Single base extension reaction system, 5 μL in total:

[0119]

[0120] Single base extension reaction procedure:

[0121]

[0122] ⑷. Electrophoresis: Take 1.2 μL of the single-base extension product obtained in step 3, add 10 μL of Hi-Di formamide and 0.5 μL of GeneScanTMSizeStandards-120LIZ, mix well, denatur...

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Abstract

The invention provides a medicolegal rapid detection kit based on 21 SNP (Single Nucleotide Polymorphism) mitochondrion loci and an application of the medicolegal rapid detection kit in individual recognition. The kit comprises an enriched primer mixture, a single-base extension primer mixture, amplification reagents, a nucleic acid purification column, an SNaPshot reaction mixed liquid, Hi-Di methanamide, GeneScan<TM>Size Standards-120LIZ, a positive control and a negative control, wherein the amplification reagents include dNTPs and rapid Taq polymerase, the positive control is DNA 9947A, and the negative control is ddH2O. The kit provided by the invention is low in cost, easy to operate, high in sensitivity, and wide in applicability, a detection result is accurate and can be acquired within 1.5-2 hours by virtue of a single-tube reaction, and precious time is saved for the individual recognition of human biological detection materials and the uncovering of cases.

Description

technical field [0001] The invention belongs to the field of forensic medicine, and in particular relates to a forensic medicine rapid detection kit based on 21 mitochondrial SNP sites and an application thereof. Background technique [0002] Forensic DNA analysis is a science and technology that uses modern DNA analysis technology to analyze the distribution and transmission rules of DNA genetic markers in the population, determine the consistency and genetic relationship of the analyzed samples, and provide evidence for the investigation and judicial trial. Its analysis objects mainly involve various body fluids (spots), tissues, hair and epidermal exfoliated cells of the human body. The main tasks of forensic DNA analysis are individual identification and paternity testing. At present, there are two main types of kits used for individual identification in forensic DNA analysis: one is based on short tandem repeat (shorttendemrepeat, STR) typing technology; the other is b...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2531/113C12Q2533/101C12Q2565/125
Inventor 严江伟杨雅冉杨猛陈婧
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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