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Fluorescent labeling composite amplification kit for 30 STR (short tandem repeat) loci of human Y chromosome and application of kit

A Y chromosome and fluorescent labeling technology, applied in the field of molecular biology, can solve the problems of low cumulative individual recognition rate, low polymorphism, and small number of loci, and achieve good amplification results, strong amplification specificity, and improved amplification. effect of possibility

Inactive Publication Date: 2016-11-23
AGCU SCIENTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

commonly used in forensic science The number of loci is small, and the cumulative individual recognition rate is low, Have been identified as having very low polymorphisms in certain populations

Method used

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  • Fluorescent labeling composite amplification kit for 30 STR (short tandem repeat) loci of human Y chromosome and application of kit
  • Fluorescent labeling composite amplification kit for 30 STR (short tandem repeat) loci of human Y chromosome and application of kit
  • Fluorescent labeling composite amplification kit for 30 STR (short tandem repeat) loci of human Y chromosome and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Screening of Y chromosome STR loci

[0043] 通过对DYS549、DYS522、DYS389I / II、DYS439、DYS447、DYS392、DYS576、DYS19、DYS388、DYS391、DYS635、DYS460、DYS456、DYS533、DYS520、DYS438、DYS527a / b、DYS449、DYS709、DYS622、DYS481、DYS437、 DYS390、DYS630、DYS448、Y_GATA_H4、DYS458、DYS607、DYS393、DYS552、DYS570、DYS385a / b、DYS593、DYS444、DYS643、DYS549、DYS557、DYS520、DYS443、DYS622、DYS587、DYS525共45个Y染色体短串联重复序列 The loci and their allelic genetic characteristics have been thoroughly studied, and 30 Y chromosome short tandem repeat loci have been screened out: DYS392, DYS389I / II, DYS447, DYS438, DYS527a / b, DYS522, DYS391, DYS456, DYS19, DYS388, DYS448, DYS385a / b, DYS481, DYS437, DYS390, DYS460, DYS533, DYS458, DYS393, Y_GATA_H4, DYS439, DYS635, DYS444, DYS643, DYS460, DYS533, DYS458, DYS393, Y_GATA_H4, DYS439, DYS635, DYS444, DYS643, DYS549, DYS557, DYS557, DYS439, DYS635, DYS444, DYS643, DYS549, DYS557. The population has low genetic polymorphism and good allele frequency distribution, which complies with ...

Embodiment 2 6

[0047] The establishment of embodiment 2 six-color fluorescent system

[0048] Fluorescence detection technology used in forensic laboratories, with the characteristics of multi-color analysis, fast and easy to use. Its main process includes: laser bombardment of a fluorophore (dye) attached to the end of the DNA, the fluorophore absorbs the laser energy, and then emits a lower energy (larger wavelength) light, and a filter is used to collect the emission of a specific wavelength range Light, using a photomultiplier tube or an electrically coupled device to collect and amplify the signal from the fluorophore, convert it into an electrical signal, and finally generate a peak map in capillary electrophoresis. Multicolor fluorescence detection technology refers to the use of different fluorescein-labeled primers for detection at each wavelength, so as to achieve the purpose of detecting composite amplification products. Considering the physical properties of fluorescein and the ...

Embodiment 3

[0049] Example 3 Arrangement of 30 Y chromosome STR loci

[0050] According to the above 30 loci, a unique locus arrangement and chemical fluorescent dye labeling method are designed: DYS392, DYS389I / II, DYS447, DYS438, DYS527a / b, DYS522 are the first group, and the fluorescent dye markers are 6 -FAM; DYS391, DYS456, DYS19, DYS388, DYS448, DYS385a / b is the second group, the fluorescent dye label is HEX; DYS481, DYS437, DYS390, DYS460, DYS533, DYS458 is the third group, the fluorescent dye label is TAMRA ; DYS393, Y_GATA_H4, DYS439, DYS635, DYS444, and DYS643 are the fourth group, and the fluorescent dye marker is ROX; DYS549, DYS557, DYS520 are the fifth group, and the fluorescent dye marker is VIG, and the loci are arranged as follows: figure 1 .

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Abstract

The invention discloses a fluorescent labeling composite amplification kit for 30 STR (short tandem repeat) loci of human Y chromosome and application of the kit. It is possible to multiple amplify 30 low-mutation-rate STR loci on Y chromosome in single reaction; by designing locus arrangements and specific primer sequences, 30 loci are divided into 5 groups, and six-color fluorescent system marks are used; the Y-STR kit is the domestically first one employing six-color fluorescent detection technology and is an STR kit detecting a highest number of loci, all the loci included are low-mutation-rate Y-STR loci, and the kit is more suitable for examining male families; the composite amplification kit is good in primer specificity, can tolerate a wide range of temperatures and is highly adaptive to subject materials.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to molecular genetics, in particular to a fluorescent labeling composite amplification kit for 30 STR loci of human Y chromosome and application thereof. Background technique [0002] Short tandem repeat locus (Short Tandem Repeat, STR) is a commonly used genetic marker. It widely exists in the genomes of prokaryotic and eukaryotic organisms, and is a kind of repeating sequence of tens to more than one hundred nucleotides composed of 2 to 6 nucleotides as repeating units. Different numbers of core sequences are arranged in tandem repeats, while the lengths are polymorphic. According to the conserved sequences at both ends, design primers, perform PCR, and then detect polymorphisms of STR genetic markers by polyacrylamide, agarose gel electrophoresis, or capillary electrophoresis. Compared with other genetic markers, STR markers have small fragments and are easy to amplify, and are more ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156
Inventor 郑卫国李科杰梅兴林郭育林
Owner AGCU SCIENTECH
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