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 operat

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

Example Embodiment

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

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

[0060] 0.1ng 9947A was used as a composite amplification template for detection. The detection steps are as follows:

[0061] ⑴. Compound amplification reaction:

[0062] Fast Taq polymerase is GoldFaster from BioDev TM Taq.

[0063] Compound amplification reaction system, total 20μL:

[0064]

[0065] Compound amplification program:

[0066]

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

[0068] ⑶. Single base extension reaction:

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

[0070] 1ng of purified composite amplification product

[0071] Single base extension primer mix 0.5μL

[0072] SNaPshot reaction mixture 1.2μL

[0073] ddH2O fill up to 5μL

[0074] Single base extension reaction program:

[0075]

[0076] ⑷. Electrophoresis: Take 1.2μL of the si...

Example Embodiment

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

[0079] Collect the following types of samples from 10 unrelated individuals: blood, semen spots, saliva, teeth, bones, nails, hair, skin, exfoliated cells, and heart, liver, spleen, lung, stomach, kidney, pancreas, brain, intestine, gallbladder Organs, a total of 190 biological specimen samples, the kit provided by the present invention and its use method are used to detect 21 SNP sites in 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; teeth 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 Extraction kit extraction;

[0083] ⑵...

Example Embodiment

[0107] Example 3

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

[0109] 0.25 pg, 0.5 pg, 0.001 ng, 0.005 ng, 0.01 ng, 0.1 ng, 1 ng, and 10 ng of 9947A were used as multiple amplification templates for detection. The detection steps are as follows:

[0110] ⑴. Compound amplification reaction:

[0111] Fast Taq polymerase is GoldFasterTM Taq from BioDev.

[0112] Compound amplification reaction system, total 20μL:

[0113]

[0114] Compound amplification program:

[0115]

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

[0117] ⑶. Single base extension reaction:

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

[0119]

[0120] Single base extension reaction program:

[0121]

[0122] ⑷. Electrophoresis: Take 1.2μL of the single-base extension product obtained in step ⑶, add 10μL of Hi-Di formamide, 0.5μL of GeneScanTMSizeStandards-120LIZ, mix well, denature at 95℃ for 3min, 4℃ quic...

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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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