Fluorescently-labeled multiplex amplification kit for 35 STR loci of human Y chromosome and application of fluorescently-labeled multiplex amplification kit

A Y chromosome and fluorescent labeling technology, applied in the field of forensic genetics, can solve the problems of narrowing the scope of investigation, unfavorable family distinction, low polymorphism of low mutation loci, etc., and achieve high sensitivity, accurate typing results, and strong specificity Effect

Pending Publication Date: 2019-12-31
安徽省公安厅物证鉴定中心 +2
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Problems solved by technology

[0004] The unique advantage of Y-STR is that all male samples of the same paternal line are of the same type, which can be used as a tool for family investigation, thereby effectively narrowing the scope of investigation. Low mutation loci are more suitable for family investigation, but low mutation loci are often less polymorphic , which is not conducive to the distinction between families

Method used

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  • Fluorescently-labeled multiplex amplification kit for 35 STR loci of human Y chromosome and application of fluorescently-labeled multiplex amplification kit
  • Fluorescently-labeled multiplex amplification kit for 35 STR loci of human Y chromosome and application of fluorescently-labeled multiplex amplification kit
  • Fluorescently-labeled multiplex amplification kit for 35 STR loci of human Y chromosome and application of fluorescently-labeled multiplex amplification kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1. Gene locus screening

[0034] 通过对DYS19、DYS385a / b、DYS389I、DYS389II、DYS390、DYS391、DYS392、DYS393、DYS437、DYS438、DYS439、DYS448、DYS456、DYS458、DYS635、Y_GATA_H4、DYS481、DYS533、DYS576、DYS643、DYS460、DYS549、DYF387S1a、 DYF387S1b、DYS449、DYS518、DYS627、DYS570、DYS527a / b、DYS447、DYS444、DYS557、DYS596、DYS446、DYS510、DYS622、DYS443、DYS587、DYS522、Y_GATA_A10、DYS520、DYS552、DYS593、DYS531、DYS459a / b、DYS508、 DYS388, DYS617, DYS645, DYS713, DYS630, DYF404S1, DYS626, DYS526a / b, a total of 59 Y-chromosomal short tandem repeat loci and their allelic genetic characteristics were thoroughly studied, and 35 Y-chromosome short tandem repeats were screened out 序列基因座:DYS392、DYS389I、DYS447、DYS389II、DYS438、DYS527a、DYS527b、DYS596、DYS391、DYS456、DYS19、DYS448、DYS385a、DYS385b、DYS549、DYS437、DYS481、DYS533、DYS390、DYS627、DYS458、DYS460、DYS393、 Y_GATA_H4, DYS439, DYS635, DYS444, DYS643, DYS557, DYS576, DYS570, DYF387S1a, DYF387S1b, DYS449, DYS518. These 35 Y chromosome short tandem repeat sequence loci have low genetic...

Embodiment 2

[0050] The method for using the fluorescent labeling compound amplification kit of 35 STR loci of human Y chromosome comprises the following steps,

[0051] A. Prepare the amplification system, the amplification system is: Reaction Mix 10.0 μL, genomic DNA X μl content is 0.125-5ng, primer mixture 5.0 μL, hot start Taq enzyme (5U / μL) 0.5 μL, sdH 2 O make up to 25.0 μL;

[0052] B. Amplification thermal cycle, the amplification program is: initial denaturation, 95°C for 2 minutes; thermal cycle, 94°C for 30 seconds, 60°C for 1min, 68°C for 1min, 30 cycles; final extension, 72°C for 20 minutes; Maintain at 4°C; among them, the amplified samples should be kept in the dark;

[0053] C. Fluorescent detection of the amplified product on the genetic analyzer, mix 12.5 μl of the sample mixture with 1 μl of the amplified product or the Allelic Ladder (Wuxi Zhongde Meilian Biotechnology Co., Ltd.) Company) mixed, denatured at 95°C for 3 minutes, ice-bathed for 3 minutes, and electroph...

Embodiment 3

[0056] Application of individual screening in the same family in embodiment 3

[0057] (1) Collection of family samples in family investigation: a total of 2 samples provided by a public security bureau.

[0058](2) Sample DNA extraction: the silica bead extraction method in the forensic DNA laboratory test specification [GA-T383-2002] was used.

[0059] (3) Using the kit of the present invention and the AGCU Database Y30 kit to perform typing detection on the above two family samples, and then compare the typing results of the two kits. Wherein, the Y-STR typing figures of the two family samples detected by the kit of the present invention are as follows: Figure 4 , Figure 5 shown.

[0060] The classification results are compared in the following table:

[0061]

[0062]

[0063] The results showed that using the AGCU Database Y30 to test the two family samples, the genotypes obtained were completely consistent, and individual identification could not be performed...

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Abstract

The invention discloses a fluorescently-labeled multiplex amplification kit for 35 STR loci of a human Y chromosome and application of the fluorescently-labeled multiplex amplification kit. The fluorescently-labeled multiplex amplification kit comprises specific primers for amplifying the 35 Y-STR loci, wherein the 35 Y-STR loci comprise 28 low-mutation Y-STR loci and 7 rapid-mutation Y-STR loci.The kit provided by the invention comprises the 28 Y-STR loci with the low mutation rate and the 7 rapid-mutation Y-STR loci with high polymorphism, and male family investigation and distinguishing between different male individuals in the same paternal line are balanced; the primers in the kit have the advantages of strong specificity, high sensitivity and accurate typing result, and can completely meet the requirements of actual case inspection, DNA database construction and paternity test; and the kit has very strong test material adaptability.

Description

technical field [0001] The invention belongs to the field of forensic genetics, and relates to a fluorescent marker compound amplification test system, in particular to a fluorescent marker compound amplification kit for 35 STR loci of human Y chromosome and its application. Background technique [0002] In the 1990s, fluorescently labeled short tandem repeat loci (short tandem repeat, STR) were first proposed and used as genetic markers for forensic evidence related research and identification. Compared with other genetic markers, STR markers have small fragments and are easy to amplify, and are more suitable for trace and degraded samples. Multiple STR loci can be simultaneously amplified, mixed samples, and DNA molecule degradation detection. Difficult biological samples such as biological samples and trace biological samples can be successfully tested and typed by DNA, so they have the advantages of fast, efficient, accurate, sensitive, and large amount of information. ...

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 陈林郑卫红王留柳张科耿学磊史明浩张雷梅兴林郑卫国
Owner 安徽省公安厅物证鉴定中心
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