Y-chromosome STR gene locus fluorescence labelled multiplex amplification kit with reinforcing resolution capability and application thereof

A Y chromosome and fluorescent labeling technology, which is applied in the field of biotechnology Y chromosome typing and identification, to achieve the effects of greatly increasing practicability and functionality, improving amplification efficiency, and improving recognition

Inactive Publication Date: 2015-06-10
NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This kind of multipurpose kit has not been reported and used yet

Method used

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  • Y-chromosome STR gene locus fluorescence labelled multiplex amplification kit with reinforcing resolution capability and application thereof
  • Y-chromosome STR gene locus fluorescence labelled multiplex amplification kit with reinforcing resolution capability and application thereof
  • Y-chromosome STR gene locus fluorescence labelled multiplex amplification kit with reinforcing resolution capability and application thereof

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment 1

[0034] Specific Example 1 Determination of Genetic Loci

[0035] 本发明通过对DYS456、DY508、A10、DYS463、DYS510、DYS464、DYS458、DYS520、DYS437、DYS439、DYS392、DYS385a / b、DYS393、DYS391、DYS390、DYS635、DYS447、DYS438、DYS389I、DYS19、GATA-H4、 The genetic polymorphism investigation of DYS389II, DYS448, DYS576, DYS570, DYS481, DYF387S1, DYS627, DYS460, DYS449, DYS533, DYS518 and other 37 loci in the male population, and finally determined DYS456, DYS576, DYS570, DYS487, , DYS458, DYS460, DYS437, DYS439, DYS392, DYS385a / b, DYS393, DYS391, DYS390, DYS635, DYS449, DYS533, DYS438, DYS389I, DYS19, GATA-H5, DYS38948, DYS2 and other genes. These loci are not only compatible with existing kits to ensure the sharing and exchange of existing DNA data, but also can improve the cumulative individual identification ability and cumulative non-parent exclusion rate of the DNA testing system, which is more in line with the technical requirements of DNA testing. In particular, the newly added 7 rapid mutation sites wi...

specific Embodiment 2

[0036] Specific Example 2 Fluorescent labeling conforms to the design of the gene locus combination scheme of the amplification system

[0037] The present invention identifies and selects fluorescent dyes, selects five fluorescent markers of blue, green, yellow, red and orange, and constructs a five-color fluorescent combination scheme. On the basis of determining the 5-color fluorescent combination scheme, through a large number of repeated experiments, the combination of loci and the type of fluorescent markers were designed. Considering the production cost and primer amplification efficiency of each locus, the 27 loci were divided into 4 groups, labeled with FAM, HEX, TAMRA, and ROX, and the molecular weight internal standard was carried out with the fluorescent dye Atto 633 in the fifth color orange. mark. After screening, a preferred fluorescent dye labeling method is finally determined: DYS456, DYS576, DYS570, DYS481, DYF387S1, DYS627 are labeled with FAM; DYS458, DYS4...

specific Embodiment 3

[0038] The present invention has the Y chromosome STR loci fluorescence labeling composite amplification kit with enhanced identification ability, and the kit includes:

[0039] 1) 2×PCR Master Mix

[0040] 2) STRtyper-27Y Primer Mix

[0041] 3) Control DNA 9948A

[0042] 4) Allelic Ladder allelic typing standard

[0043] 5) Size-500 orange fluorescent molecular weight internal standard

[0044] 6) Spectral calibration standard

[0045] The above PCR Master Mix includes: DMSO 10mM, Tris-buffer 125mM, Potassium Chloride 125mM, Ammonium Sulfate 65mM, etc., which can achieve compatible amplification of various common test materials in the market.

[0046] The above STRtyper-27Y Primer Mix includes all primers for amplifying 27 loci (see Table 2 for concentration), deoxynucleotide triphosphates (dNTPs) 7.5mM, Taq enzyme 5U / μl, magnesium chloride 7.5Mm, BSA2.5mg / ml Wait.

[0047] The above-mentioned positive control substance is purchased human genomic DNA.

[0048] The Alle...

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Abstract

The invention discloses a Y-chromosome STR gene locus fluorescence labelled multiplex amplification kit with reinforcing resolution capability. The kit comprises the following specific oligonucleotides amplification primer pairs of 27 Y-chromosome STR gene loci: DYS456, DYS458, DYS437, DYS439, DYS392, DYS385a/b, DYS393, DYS391, DYS390, DYS635, DYS438, DYS389I, DYS19, GATA-H4, DYS389II and DYS448, wherein the kit is characterized by additionally comprising at least one of the specific oligonucleotides amplification primer pairs of 10 Y-chromosome STR gene loci: DYS576, DYS570, DYS481, DYS627, DYS460, DYS449, DYS533, DYS518 and DYF387S1. The number of the detection loci of the kit is greater than that of the market similar product both at home and abroad, so that the discrimination power and the combined power of exclusion of the system are greatly improved and the individual discriminability is generally improved. Simultaneously the invention creatively comprises 7 rapid mutation sites, so that the discrimination capacity of the close relative males can be increased, the practicability and the functionality of the kit are greatly increased, and a vicious cycle of blindly increasing the similar detection sites by the domestic kit developers is avoided.

Description

technical field [0001] The invention belongs to the field of biotechnology Y chromosome typing and identification. The invention relates to a PCR amplification kit, in particular to a fluorescent labeling composite amplification kit for simultaneously detecting 27 Y chromosome STR loci in a single tube and the reagent The application of boxes in the field of forensic identification. Background technique [0002] Short tandem repeat (short tandem repeat, STR) is a type of DNA sequence with length polymorphism formed by tandem repetition of 2-6 bases in the human genome as the core unit. The number of core units varies and the number of repetitions is different. The genetic polymorphisms that make up the STR. STRs are widely distributed and numerous, accounting for about 10% of the human genome, and contain a huge amount of information. Different sequences can generate hundreds of millions of genotype combinations, and the frequency of each combination in the population is ve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/68
Inventor 李政刘亚举张兹均金海英吴勇宋丹璐
Owner NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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