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Kit for synchronously detecting 124 micro haplotype gene loci on basis of next generation sequencing technology and special primer pair combination thereof

A technique of primer pair and haplotype applied in the field of forensic science

Active Publication Date: 2018-10-09
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Thermo Fisher in the United States is testing and evaluating the next-generation sequencing kit containing micro-haplotypes, namely EarlyEvaluation Precision ID Mixture ID Panel, but it only contains 36 micro-haplotype loci, and the data analysis program supporting the kit can only analyze the sequencing data of the inherent loci in the kit, which has certain limitations

Method used

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  • Kit for synchronously detecting 124 micro haplotype gene loci on basis of next generation sequencing technology and special primer pair combination thereof
  • Kit for synchronously detecting 124 micro haplotype gene loci on basis of next generation sequencing technology and special primer pair combination thereof
  • Kit for synchronously detecting 124 micro haplotype gene loci on basis of next generation sequencing technology and special primer pair combination thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Preparation of a kit for detecting micro-haplotype loci based on next-generation sequencing technology

[0048] 1. Preparation of primer pair combinations

[0049] The primer set consisted of 248 primers for the detection of 124 micro-haplotype loci. The name of each micro-haplotype locus, the name of the primer corresponding to the amplified micro-haplotype locus, the nucleotide sequence of the primer, and the allelic genotype are shown in Table 1. The length and nucleotide sequence of the PCR amplification products of the primers corresponding to each micro-haplotype locus in the hg19 human reference genome are shown in Table 2.

[0050] Table 1

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[0052]

[0053]

[0054]

[0055] Table 2

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[0057]

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[0059]

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[0061]

[0062] 2. Preparation of kits for detecting micro-haplotype loci based on next-generation sequencing technology

[0063] A kit for detecting micro-haplotype loci based on nex...

Embodiment 2

[0064] Example 2. Application of the kit for detecting micro-haplotype loci based on next-generation sequencing technology

[0065] 1. DNA sample preparation

[0066] according to The instruction manual of M48 DNA Manual Kit (200) is to extract the genomic DNA of a female oral epithelial cell sample. Take 1 μL DNA template and use Qubit TM dsDNA HS Assay Kit, 500 assays for sample DNA quantification, and then diluted with ultrapure water to obtain a genomic DNA aqueous solution of female oral epithelial cells at a concentration of 0.2ng / μL, referred to as female genomic DNA solution.

[0067] according to The instruction manual of M48 DNA Manual Kit (200) is to extract the genomic DNA of a male blood card sample. Take 1 μL DNA template and use Qubit TM dsDNA HS Assay Kit, 500 assays for sample DNA quantification, and then diluted with ultrapure water to obtain a male blood card genomic DNA aqueous solution with a concentration of 0.2ng / μL, referred to as male genomi...

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Abstract

The invention discloses a kit for synchronously detecting 124 micro haplotype gene loci on the basis of a next generation sequencing technology and a special primer pair combination thereof. The primer pair combination provided by the invention is composed of 248 molecules shown as sequences 1-248 in a sequence table. Experiments prove that the kit for synchronously detecting 124 micro haplotype gene loci on the basis of the next generation sequencing technology, provided by the invention, is used for detecting 124 micro haplotype gene loci of human genome DNA, and the typing result is accurate. The kit provided by the invention has a significant application value.

Description

technical field [0001] The invention relates to the technical field of forensic medicine, in particular to a kit for synchronously detecting 124 micro-haplotype loci based on next-generation sequencing technology and a special primer pair combination thereof. Background technique [0002] Forensic DNA technology based on forensic science and genetics has been widely used in the fields of criminal case investigation and evidence collection, combating trafficking in women and children, identification of disaster victims and missing persons, identification of civil paternity relationship, etc. STR typing based on the capillary electrophoresis platform is the gold standard for individual identification in the field of forensic science today. After nearly 30 years of accumulation, the field of forensic DNA has been able to solve the DNA testing problems of routine biological samples such as blood, saliva, semen, bones, and muscles, but the DNA testing and typing of two or more pe...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q1/6888C12Q2600/16C12Q2600/172C12Q2535/122
Inventor 王乐饶旼张驰季安全李彩霞严安心宋娇娇叶健
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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