Composite amplification system based on Y-STR locus and specific primer combination thereof

A compound amplification system and primer combination technology, applied in the field of forensic science, can solve the problems that different individuals cannot be distinguished well, the mutation rate of Y-STR loci is not high, and the application of Y-STR loci is limited.

Active Publication Date: 2018-11-23
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

[0004] However, with the continuous development of society, more complicated cases have brought greater challenges to the public security organs
Especially in some sexual assault cases, on-site DNA samples are highly mixed, and sometimes only the Y-STR typing of the suspect can be obtained
In the identification of a large number of male individuals, the multiple amplification system of the existing Y-STR loci has shown shortcomings in terms of the number of loci and the mutation rate of the loci: on the one hand, the existing Y-STR loci Single-copy Y-STR loci accounted for the majority in the complex amplification system, and the only individual multi-copy Y-STR loci had only 2 copies, and the detected polymorphisms were very limited; on the other hand, the commonly used Y-STR loci had only 2 copies. The mutation rate of STR loci is not high, about 1×10 -3 Even lower, which leads to the inability to distinguish between different individuals in the same family, which limits the application of the Y-STR locus in individual identification to a certain extent

Method used

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  • Composite amplification system based on Y-STR locus and specific primer combination thereof
  • Composite amplification system based on Y-STR locus and specific primer combination thereof
  • Composite amplification system based on Y-STR locus and specific primer combination thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0122] Embodiment 1, the preparation of the compound amplification system based on Y-STR locus

[0123] 1. Screening of Y-STR loci

[0124] The inventor of the present invention obtains a large amount of candidate multi-copy Y-STR loci by consulting the literature; Then, using the research results of the Human Genome Project, using the reported The primer information of each locus was retrieved to obtain the corresponding sequence; the sequence was retrieved using BLAST (URL: https: / / blast.ncbi.nlm.nih.gov / Blast.cgi), and the existing multi-copy Y - Sequence information of the core and flanking regions of the STR locus, allele length and distribution, etc. were sorted out and systematically studied, focusing on multi-copy Y-STR loci with a core sequence of 4-5 base repeats. Based on technical indicators such as copy number, mutation rate, repeat unit characteristics, and sequence structure complexity of each locus, 35 candidate multi-copy Y-STR loci were initially screened ou...

Embodiment 2

[0135] Embodiment 2, the application of the compound amplification system based on Y-STR locus

[0136] The sample to be tested is sample 1 or sample 2:

[0137] Sample 1, 1ng of human (known as male) genomic DNA;

[0138] Sample 2: Take a blood collection card from a person (known to be a male), and use a 0.5mm hand-held punch to punch out a disc.

[0139] The blood collection card of the person (known as a male) was provided by the Daily DNA Database Construction Institute of the Physical Evidence Identification Center of the Ministry of Public Security.

[0140] 1. Take 10 μL of the compound amplification system based on the Y-STR locus prepared in Step 3 of Example 1, add the sample to be tested for PCR amplification, and obtain the PCR amplification product.

[0141] Reaction program: pre-denaturation at 95°C for 11 min; denaturation at 94°C for 30 s, annealing at 59°C for 2 min, extension at 72°C for 1 min, 28 cycles; extension at 60°C for 60 min; storage at 4°C.

[0...

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Abstract

The invention discloses a composite amplification system based on a Y-STR locus and a specific primer combination thereof. The primer combination provided by the invention is formed by 30 kinds of DNAmolecules shown as sequence 1 to sequence 30 in a sequence table. The primer combination is used for building the composite amplification system based on the Y-STR locus; then, male individual STR typing is performed; the detected polymorphism is high; the population screening and individual recognition capability of a Y chromosome genetic marker can be enhanced; particularly important identification values are realized on the distant relatives of the same family. Therefore when the primer combination provided by the invention is used for sample screening, the cost is low; the efficiency is high; important application values are realized.

Description

technical field [0001] The present invention relates to the technical field of forensic medicine, in particular to a complex amplification system based on Y-STR loci and its special primer combination, especially to a multi-copy Y-STR locus based on 13 multi-copy Y-STR loci, 1 single-copy locus DYS391 and 1 A multiplex amplification system of a sex identification locus Amelogenin and its special primer combination. Background technique [0002] Short tandem repeats (short tandem repeats, STRs) belong to the second generation of genetic markers. Due to their small repeat units, high mutation rate, and wide distribution in the genome, they have been widely used in the fields of population genetics and forensic science. In the human genome, there is one STR site about every 15-20kb, and they account for 10% of the total human genome. These STRs usually consist of tandem repeating units of 2-6 bases. Due to the difference in repeating units and repeating times, they show large ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2525/151
Inventor 尚蕾孙辉杨帆莫晓婷赵兴春李万水叶健
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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