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Individual recognition system based on next generation sequencing and kit and application of kit

A second-generation sequencing and kit technology, applied in the field of genes, can solve the problems of high price, long PCR product length, insufficient compatibility, etc., and achieve the effect of reducing operation steps, high sensitivity, and shortening experimental time.

Active Publication Date: 2018-09-11
BGI FORENSIC TECH (SHENZHEN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Illumina MiSeq FGx based on next-generation sequencing technology is expensive, and some recognition sites are poorly polymorphic in the Chinese population, which is not suitable for DNA typing of the Chinese population
At the same time, the platform compatibility of Illumina MiSeq FGx is not enough and some PCR products are long in length, making it difficult to handle old and highly degraded samples

Method used

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  • Individual recognition system based on next generation sequencing and kit and application of kit
  • Individual recognition system based on next generation sequencing and kit and application of kit
  • Individual recognition system based on next generation sequencing and kit and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] This embodiment provides an individual identification system based on next-generation sequencing. The individual identification system genotypes a male sample and detects the blood type.

[0088]The blood was donated by male volunteers, and the serological test showed blood type B. The DNA template was extracted with the chelex-100 method, and the primers were synthesized by a company and mixed according to a certain ratio to form a primer mixture. Take the primer mixture, DNA polymerase Mix, etc., and configure the reaction system according to Table 8. Set up a thermal cycler (ABI Veriti PCR instrument) according to the reaction conditions in Table 9, and put the PCR reaction tube into the instrument for PCR amplification. After the amplification reaction is completed, take out the reaction tube and use Ultra TM DNALibrary Prep Kit for Library preparation kits to prepare DNA libraries. Take the PCR purified product, according to Ultra TM DNA Library Prep Ki...

Embodiment 2

[0101] This embodiment provides an individual identification system based on next-generation sequencing. The individual identification system genotypes a female sample and detects the blood type.

[0102] The blood was donated by female volunteers, and it was type A blood by serological test. Template DNA was extracted by chelex-100 method. The amplification reaction was carried out on an ABI 9700 thermal cycler with Ultra TM DNA Library Prep Kit for The library preparation kit was used to build the library, and the Illumina Hiseq 4000 was used for sequencing.

[0103] Among them, the autosomal typing results of female samples are shown in Table 12, and the detection results of blood group-related loci of female samples are shown in Table 13.

[0104] Table 12. Experimental data and typing results of autosomes in female samples

[0105] site name

[0106] Table 13. Detection results of blood group-related loci

[0107] Allele

[0108] According to ...

Embodiment 3

[0110] This embodiment provides an individual identification system based on next-generation sequencing. The individual identification system performs genotype analysis on two families. The two families are positive family and negative family.

[0111] Blood was donated by volunteers, and DNA templates were extracted using the chelex-100 method. The amplification reaction was carried out on an ABI 9700 thermal cycler with Ultra TM DNA Library Prep Kit for The library preparation kit was used to build the library, and the Illumina Hiseq 4000 was used for sequencing.

[0112] The autosomal typing results of positive families are shown in Table 14, and the results of autosomal typing of negative families are shown in Table 15.

[0113] Table 14. Definite family experimental data and typing results

[0114]

[0115]

[0116]

[0117] Table 15. Experimental data and typing results of negative families

[0118]

[0119]

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PUM

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Abstract

The invention discloses an individual recognition system based on next generation sequencing and a kit and application of the kit. The individual recognition system comprises a primer sequence designed at 112 STR sites and 318 SNP sites, wherein the 112 STR sites include 53 autosome STR sites, 23 X chromosome STR sites, 35 Y chromosome STR sites and a gender determination site AMEL; the 318 SNP sites include 186 autosome SNP sites, 69 X chromosome SNP sites, 57 Y chromosome SNP sites and 6 blood group phenotype related SNP sites. The individual recognition system can obtain second-generation or three-generation genetic marker information on autosome, X chromosome and Y chromosome at the same time only through once amplification; thus, operation steps are reduced, experiment time is shortened, and the individual recognition system is suitable for DNA genotype detection of people in different countries and different areas all over the world.

Description

technical field [0001] The invention relates to the field of gene technology, in particular to an individual identification system based on next-generation sequencing, a kit and its application. Background technique [0002] Short tandem repeats (Short Tandom Repeat, STR), also known as microsatellite DNA or simple sequence repeats (Simple Sequence Repeats, SSR), are a type of repetitive sequence widely present in the human genome, which consists of 2-6 bases The core sequence is arranged in tandem repeats. STR is widely used in individual identification, paternity identification and population genetics research because of its large number, wide distribution, high genetic polymorphism, and simple detection method in the human genome. [0003] SNP (Single Nucleotide Polymorphism, single nucleotide polymorphism) refers to changes such as transitions, transversions, insertions, and deletions at a specific nucleotide position in DNA in the genome. As the third generation of ge...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/156
Inventor 李生斌王泳钦杨慧龚雨晴
Owner BGI FORENSIC TECH (SHENZHEN) CO LTD
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