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Multiplex amplification detection system and method including 13 rapidly-mutating Y-STR loci and application

A technology of compound amplification and detection method, which is applied in the field of compound amplification detection system, can solve the problems of not providing system details, related primer design defects, etc., and achieve a well-balanced effect

Active Publication Date: 2020-07-31
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

[0009] The second is that individual loci in the system contain multiple copies, and the design of related primers has certain defects
[0010] In the research of domestic Chen Yuling et al. (2017), the self-established rapid mutation Y-STR system was used, but the details of the system were not provided

Method used

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  • Multiplex amplification detection system and method including 13 rapidly-mutating Y-STR loci and application
  • Multiplex amplification detection system and method including 13 rapidly-mutating Y-STR loci and application
  • Multiplex amplification detection system and method including 13 rapidly-mutating Y-STR loci and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Composite amplification detection system containing 13 rapidly mutating Y-STR loci: including 13 loci and fluorescent labeling dyes corresponding to the primers of the 13 loci.

[0068] The structures of five fluorescent dyes are shown in Table 1:

[0069] Table 1

[0070]

[0071] An orange fluorescent dye was used as an internal standard dye.

[0072] The loci labeled with different fluorescent dyes are shown in Table 2:

[0073] Table 2

[0074]

[0075]

[0076] The specific amplification primers corresponding to the 13 loci are as shown in Table 3, wherein PCRU represents the upstream primer, and PCRL acts as the downstream primer.

[0077] table 3

[0078]

[0079] It can be seen from Table 3 that: yellow fluorescent DYF403S1a corresponds to one primer pair; red fluorescent DYF403S1b1 and DYF403S1b2 correspond to another primer pair.

[0080] DYS526 has only one pair of primers, and DYS526a and DYS526b can be amplified simultaneously.

[0081] In...

Embodiment 2

[0084] A multiplexed amplification assay comprising 13 rapidly mutating Y-STR loci.

[0085] (1) Construction of fluorescent system: 5 different fluorescent dyes shown in Table 4 are used to label the amplification primers of 13 rapidly mutating Y-STR loci, and the amplification primers of 13 rapidly mutating Y-STR loci are as follows: The nucleotide sequence of SEQ ID No.1 to SEQ ID No.28 in the sequence listing (shown in Table 3);

[0086] (2) Construction of a multiplex amplification system: use the fluorescent dye-labeled amplification primers obtained in step (1) to amplify the DNA to be detected to obtain an amplification product;

[0087] 2.1. Prepare the PCR reaction mixture, as shown in Table 4:

[0088] Table 4

[0089]

[0090] The concentrations of fluorescent amplification primers were:

[0091] DYF387S1: The concentrations of upstream primers and downstream primers are 0.04 μM,

[0092] DYF399S1: the concentrations of upstream primers and downstream primer...

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PUM

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Abstract

The invention discloses a multiplex amplification detection system and method including 13 rapidly-mutating Y-STR loci and application. The multiplex amplification detection system includes the 13 loci and fluorescent labeling dyes corresponding to primers of the 13 loci. The invention develops a set of multiplex amplification detection system including the 13 rapidly-mutating Y-STR loci, and a five-color fluorescent technique (blue FAM, green HEX, yellow TAMRA, red ROX and orange) is selected to perform multiplex amplification, so that good amplification: a peak pattern is sharp, no blended bands are generated and the balance is good, of a DNA sample can be realized. The system can be applied to refined identification in male family screening, and provides a technical support for investigation and case handling.

Description

technical field [0001] The invention relates to the technical field of genetic markers. Specifically, it is a compound amplification detection system, method and application comprising 13 rapidly mutated Y-STR loci. Background technique [0002] Y-STR genetic marker is an effective means of inspection and identification in the field of forensic science, and it is an important supplement to autosomal STR and mitochondrial genetic markers. Since the Y chromosome is unique to male individuals, most of it (more than 95%) is a non-recombination region, and Y-STR presents the characteristics of paternal inheritance, which plays an important role in the identification of paternal genetic relationship and the examination of male individuals with mixed spots. At the same time, using the relatively conservative characteristics of Y-STR in a family, Y-STR typing tests can be carried out on families in key areas, and family investigations can be carried out, thereby assisting investiga...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2531/113C12Q2525/151C12Q2563/107C12Q2565/125
Inventor 尚蕾丁光树莫晓婷白雪马温华李万水孙辉
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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