Fluorescent multiplex amplification kit for detecting human Y-STR loci and application of fluorescence multiplex amplification kit

A Y-STR, compound amplification technology, applied in the field of molecular genetics, to achieve the effect of rapid amplification, simple operation method and good balance

Pending Publication Date: 2021-04-06
AGCU SCIENTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved: In order to overcome the shortcomings of the existing technology and adapt to the complexity of the current case scene, provide a reagent kit with strong anti-interference ability and high sensitivity, which can provide clues and evidence for solving the case and meet the needs of fast and efficient criminal investigation. , the invention provides a fluorescent multiplex amplification kit for detecting human Y-STR loci and its application

Method used

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  • Fluorescent multiplex amplification kit for detecting human Y-STR loci and application of fluorescence multiplex amplification kit
  • Fluorescent multiplex amplification kit for detecting human Y-STR loci and application of fluorescence multiplex amplification kit
  • Fluorescent multiplex amplification kit for detecting human Y-STR loci and application of fluorescence multiplex amplification kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1. Screening of 50 very low mutation Y-STR loci.

[0027] 通过对DYS618、DYS640、DYS497、DYS567、DYS590、DYS579、DYF390S1、DYS615、DYS621、DYF381S2、DYF380S1、DYS545、DYS619、DYS559、DYS595、DYS536、DYS478、DYS577、DYF405S1、DYS502、DYS642、DYS625、DYF382S1、DYS388、 DYS426、DYS435、DYS436、DYS581、DYS550、DYS453、DYS494、DYS584、DYS641、DYS477、DYS480、DYS575、DYS474、DYS588、DYS499、DYS448、DYS530、DYS538、DYS512、DYS558、DYS583、DYS434、DYS485、DYS580、DYS624、 DYS491、DYS573、DYS473、DYS571、DYS475、DYS470、DYS634、DYS455、DYS613、DYS488、DYS472、DYS489、DYS454、DYS476、DYS554、DYS561、DYS521、DYS574、DYS631、DYS525和DYS578共70个Y染色体STR基因座的 基因多态性和突变率的研究,最后筛选出DYS618、DYS640、DYS497、DYS567、DYS590、DYF390S1、DYS502、DYS615、DYS621、DYF381S2、DYF380S1、DYS545、DYS559、DYS595、DYS536、DYS478、DYS577、DYF405S1、DYS642 、DYS625、DYF382S1、DYS436、DYS550、DYS453、DYS494、DYS641、DYS477、DYS480、DYS575、DYS474、DYS588、DYS499、DYS530、DYS538、DYS512、DYS558、DYS485、DYS580、DYS624、DYS491、DYS573、DYS473、DYS571、DYS475 , DYS470, DYS634, DYS455, DYS613, DYS488, DYS472, a total of 50 very...

Embodiment 2

[0031] 1. PCR reaction buffer and amplification program adjustment

[0032] (1) The buffer of the PCR reaction (ie, A-Reaction Mix) contains various ions and dNTP mixtures necessary for the amplification reaction, and some specific PCR enhancers can also be added according to the needs of PCR. However, in order to improve the amplification specificity, amplification efficiency and anti-interference ability of the PCR reaction, it is necessary to optimize the concentration of the above components.

[0033] The optimized concentration ranges of the components contained in the PCR reaction buffer in the kit of the present invention are as follows.

[0034]

[0035] (2) The PCR amplification procedure is different from ordinary PCR in that no extension step is required. The specific amplification procedure is:

[0036]

[0037] 2. Amplification reaction system and data analysis.

[0038] The components and content of the amplification system in this kit are shown in the t...

Embodiment 3

[0041] Example 3 Different Family Male Individual Sample Test

[0042] Two male individual samples from different families were selected, and the amplification product was analyzed according to the method provided in Example 4. The results are as follows:

[0043]

[0044]

[0045] The results showed that there was only 4 steps difference between the two samples at Y37, so they might not be in the same family. The Y50 test showed that the typing of 50 loci was the same, and they belonged to the same family. Highly mutated loci may lead to false positive exclusions. The Y50 hypomutation locus could serve as strong support. It also proves that the kit has the ability to distinguish families. The kit is fully compatible with the current PCR-capillary electrophoresis platform popularized in forensic DNA laboratories. Amplify.

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Abstract

The invention discloses a fluorescence multiplex amplification kit for detecting human Y-STR loci and application of the fluorescence multiplex amplification kit. The kit comprises specific amplification primers of 50 pairs of Y-STR loci. Compared with the prior art, the kit disclosed by the invention has the following advantages that: (1) 50 pairs of STR loci with extremely low mutation on human Y chromosome are adopted, rapid amplification, good balance and high sensitivity are realized, and the requirements of individual identification, paternity test, DNA family tree construction, family investigation and the like of male criminals can be met; and (2) the kit can realize rapid batch detection of 90 samples within 2 hours, and 50 Y-STR loci can be detected in each sample. The whole operation and result analysis method is completely compatible with a PCR- capillary electrophoresis platform popularized in a forensic DNA laboratory at present, and the operation method is simple and easy to master.

Description

technical field [0001] The invention belongs to the technical field of molecular genetics, and relates to a composite amplification detection kit for detecting an extremely low mutation Y-STR locus, in particular to a fluorescent composite amplification kit for detecting a human Y-STR locus and an application thereof. Background technique [0002] The Y chromosome STR genetic marker refers to the short tandem repeat sequence that exists in the non-recombination region of the human Y chromosome. Y chromosome STR genetic markers have three major characteristics of male-specific, paternal inheritance and haplotype inheritance. Combining its uniqueness with the advantages of STR locus typing detection, it can be used for forensic individual identification, paternity identification and DNA family tree construction, etc. . [0003] In forensic science, a locus with a mutation rate higher than 1% is called a rapidly mutating locus (Rapidly Mutating Y-STR). The rapid mutation locu...

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 李娟梅兴林史明皓郑卫国
Owner AGCU SCIENTECH
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