Microhaplotype genetic marker for forensic detection and kit thereof

A genetic marker and kit technology, applied in the field of forensic medicine, can solve the problems of inability to analyze, high quality requirements, unbalanced allele amplification, etc.

Inactive Publication Date: 2018-09-07
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

[0003] Early DNA analysis was mainly restriction fragment length polymorphism (RLFP). The technical principle was to detect the size of a specific DNA fragment formed after restriction endonuclease digestion of DNA. For example, point mutations, such as new creation and removal of restriction sites, and reorganization of a piece of DNA, such as insertions and deletions that cause changes in the length of restriction sites, can lead to the generation of RFLP, but this method has a great impact on the quality of DNA. The requirements are high, so it is impossible to analyze the degraded or trace amount of DNA; with the completion of the polymerase chain reaction (PCR) and the human genome project, the short tandem repeat sequence (STR) has become the second generation of genetic markers widely used in forensic science, STR has the characteristics of high polymorphism, and the length of the amplified allele fragment is generally less than 400bp. It is a very mature forensic genetic marker at present. For example, Chinese patent 201610922569.2 discloses a kit for rapid identification of forensic autosomal STR typing. It includes a box body and a flip-type box cover connected with the box body. A paper tray is plac...

Method used

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  • Microhaplotype genetic marker for forensic detection and kit thereof
  • Microhaplotype genetic marker for forensic detection and kit thereof
  • Microhaplotype genetic marker for forensic detection and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 1

[0070] Example 1 A group of genetic marker combinations

[0071] This embodiment lists a group of genetic marker combinations, the genetic marker combinations include 22 micro-haplotypes respectively located on 22 human autosomes, the SNP loci of the 22 micro-haplotypes are composed and The chromosome number where it is located is: the SNP site is named after the rs number in the SNP database of the National Center for Biotechnology Information:

[0072] Micro-haplotype 1: rs6688242, rs2796542, rs6688263 and rs11206895, a total of 4 SNP sites, on Chr1;

[0073] Micro-haplotype 2: rs3771843, rs3821317, rs17640500, rs10194954, rs10207441, rs3771844, a total of 6 SNP sites on Chr2;

[0074] Micro-haplotype 3: rs496999, rs497931, rs623776, a total of 3 SNP sites, on Chr3;

[0075] Micro-haplotype 4: rs10028632, rs9995529, rs10028725, rs10028726, rs10028792, a total of 5 SNP sites on Chr4;

[0076] Micro-haplotype 5: rs7722440, rs6555369, rs6555370, rs11420883, a total of 4 SNP ...

Embodiment 2

[0094] Embodiment 2 A kind of compound amplification system

[0095] In this embodiment, the complex amplification system includes: 22 sets of primers for amplifying the above-mentioned 21 micro-haplotypes and Amelogenin loci and the concentration of each set of primers in the amplification system.

[0096] In this embodiment, 22 sets of primers and their primer concentrations as shown in Table 3 are used to simultaneously multiplex and amplify the 21 micro-haplotypes and Amelogenin loci in Example 1, so that all 21 micro-haplotypes and Amelogenin loci All were amplified and all were amplified with relatively consistent amplification efficiencies.

[0097] table 3

[0098]

Embodiment 4

[0115] Example 4 Frequency Calculation of Micro-haplotype Markers

[0116] Since the above micro-haplotypes are independently developed and designed, there is no corresponding frequency information, so the frequencies of these markers need to be calculated first.

[0117] The calculation method is:

[0118] (1) DNA extraction: DNeasy Blood & Tissue Kit was used to extract the whole genome DNA of 50 unrelated individuals.

[0119] (2) Multiplex PCR amplification: The multiplex amplification system constructed by the present invention is used to perform multiplex PCR amplification on the extracted DNA.

[0120] (3) Library construction and next-generation sequencing: Purify the PCR products, and use the KAPA LTP Library Preparation Kit kit to build a library of the purified PCR products. After the library inspection is qualified, use the MiSeq sequencer for sequencing.

[0121] (4) Next-generation sequencing data analysis: perform quality control on the fastq files obtained by...

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Abstract

The invention belongs to the field of forensic medicine, and discloses a microhaplotype genetic marker for forensic detection and a kit thereof. The invention provides a genetic marker combination consisting of 21 microhaplotypes located on autosomal chromosomes. The genetic marker combination can be effectively applied to forensic detection. Compared with the traditional detection kit, the microhaplotype genetic marker for forensic detection and the kit thereof provided by the invention have high sensitivity, accurate detection result and high applicability in individual identification, can realize individual identification of human biological samples in a relatively short period of time, and wins valuable time for solving of forensic cases.

Description

technical field [0001] The invention belongs to the field of forensic science, and in particular relates to a micro-haplotype genetic marker and a frequency kit for forensic detection. Background technique [0002] Forensic DNA technology based on the background of forensic science and genetics has been widely used in the fields of criminal case investigation, evidence collection, combating trafficking in women and children, identification of disaster victims and missing persons, identification of civil paternity relationship, etc. The development and progress of forensic genetics History is the history of the discovery, research and detection of new genetic markers. The development of polymorphic loci that can be used in forensic medicine has always been a hotspot and basic issue in forensic genetics research. [0003] Early DNA analysis was mainly restriction fragment length polymorphism (RLFP). The technical principle was to detect the size of a specific DNA fragment form...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 严江伟赵晶杨雅冉刘志勇陈曼陈彤
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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