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Accurate human DNA typing method, reagent and application

A DNA typing and precise technology, applied in the direction of recombinant DNA technology, DNA / RNA fragments, biochemical equipment and methods, etc., can solve problems such as maternal traceability and database construction defects, and achieve high accuracy, high individual recognition, The effect of high adaptability to inspection materials

Pending Publication Date: 2020-03-06
BGI FORENSIC TECH (SHENZHEN) CO LTD
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AI Technical Summary

Problems solved by technology

NGS-based detection technologies, such as Illumina FGx, still have certain shortcomings for matrilineal traceability and database construction and investigation of gender-based crimes

Method used

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  • Accurate human DNA typing method, reagent and application
  • Accurate human DNA typing method, reagent and application
  • Accurate human DNA typing method, reagent and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0041] 1. Site selection and primer design

[0042] In this case, 143 STRs and 188 SNPs were finally screened out through statistical research on the polymorphism and mutation rate of STR sites, SNP sites, and mitochondrial hypervariable regions, combined with the STR sites required by the public security database. , two segments of mitochondrial hypervariable regions and Amel enamel gene.

[0043] Specific primers were designed on the flanks of 143 STRs, 188 SNPs, two mitochondrial hypervariable regions and Amel enamel gene. The primers were designed using PrimerPrimier5 and Oligo7 software, and the annealing temperature of each primer was around 58°C, which could not produce primer dimers or other non-specific products caused by mismatching, and the length of the amplified product was between 150-400bp. The sequences of the final primers in this example are shown in Table 1.

[0044] Table 1 Detection sites and primers

[0045]

[0046]

[0047]

[0048]

[00...

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Abstract

The application discloses an accurate human DNA typing method, a reagent and application. According to the accurate human DNA typing method, a short tandem repeat sequence, polynucleotide polymorphicsites covering a whole genome, a mitochondrial DNA hypervariable region I, a mitochondrial DNA hypervariable region II and an Amel enamel gene are detected simultaneously to obtain accurate typing andbase sequences of all the sites, thereby realizing accurate typing of human DNAs. The disclosed method having high resolution ratio is capable of carrying out efficient and accurate individual recognition and has the extremely high individual recognition capacity for difficult detection materials and highly-degraded detection materials; the non-parent exclusion rate is larger than 99.999999% in paternity identification and the accuracy is high; the compatibility is high, the existing individual recognition detection kit can be covered, and the method can be used for analyzing race groups in different regions or countries by combining human genome data. Besides, the method disclosed by the invention is relatively high in adaptability of detection materials and the human DNA extracted by various experimental methods can be detected.

Description

technical field [0001] This application relates to the field of human DNA typing detection methods, in particular to a method, reagent and application for precise human DNA typing. Background technique [0002] Short tandem repeats (short tandem repeats, STR), also known as microsatellite DNA or simple sequence repeats (simple sequence repeats, SSR), are currently the most widely used length polymorphic genetic markers in forensic evidence identification. The repeat unit is short, only 2-6bp, and its length polymorphism comes from the individual differences in the copy number of the repeat unit. [0003] Within the scope of the human genome, if any base mutation causes two bases to appear at a specific nucleotide position, the least one of which has a frequency of not less than 1% in the population, a single nucleotide polymorphism (SNP) is formed ( single nucleotide polymorphisms, SNPs). Due to the rich content of SNP and its high genetic stability, its application in for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 李生斌苗鑫垚常辽于慧云
Owner BGI FORENSIC TECH (SHENZHEN) CO LTD
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