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Multi-color fluorescence multiplex amplification kit for amplifying STR gene locuses of human Y chromosome and application of kit

A Y-chromosome and compound amplification technology, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of low individual identification and individual identification that cannot be unique, and achieve high individual identification and multiple The effect of high morphism and low mutation rate

Inactive Publication Date: 2017-03-22
AGCU SCIENTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with autosomal STR loci, Y-STR haplotypes have lower individual recognition ability in individual identification, and cannot be used for unique individual identification, and can only be used as a tool to assist public security criminal investigation investigations

Method used

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  • Multi-color fluorescence multiplex amplification kit for amplifying STR gene locuses of human Y chromosome and application of kit
  • Multi-color fluorescence multiplex amplification kit for amplifying STR gene locuses of human Y chromosome and application of kit
  • Multi-color fluorescence multiplex amplification kit for amplifying STR gene locuses of human Y chromosome and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Screening of Y chromosome STR locus

[0030]A total of 2165 male blood samples from 1008 unrelated male individuals and 1124 father-son pairs were collected from the Chinese Han population, and the genetic polymorphisms and mutations of 48 Y-STR loci were tested and analyzed in the Chinese Han population. 所有测试基因座如下:DYS459a / b、DYS643、DYS522、DYS389I / II、DYS447、DYS576、DYS19、DYS388、DYS391、DYS635、DYS456、DYS533、DYS520、DYS622、DYS481、DYS437、DYS390、DYS630、DYS448、GATA_H4、 DYS458、DYS393、DYS552、DYS570、DYS385a / b、DYS593、DYS444、DYS438、DYS527a / b、DYS392、DYS460、DYS449、DYS549、DYS439、DYS587、DYS446、DYS518、DYS627、DYS557、DYF387S1a / b、DYS510、DYS443、 GATA_A10, DYS531, finally screened out 30 Y chromosome STR loci with polymorphism greater than 0.6 and mutation rate less than 5‰.

[0031] The statistics of polymorphism and mutation rate information of 48 Y-STR loci are shown in Table 3:

[0032] Table 3 Statistics of polymorphism and mutation rate information of 48 Y-STR loci

[0033] ...

Embodiment 2

[0039] Example 2 Y chromosome STR locus arrangement

[0040] Kit locus assignments are performed by combining primer design and the allelic range of each locus. The following points need to be paid attention to during the design process: a. Choose fluorescent dyes with a small degree of overlap; b. Arrange as many loci as possible for fluorescent dyes with high efficiency, and place fewer loci for those with low efficiency; c. Low efficiency The loci need to be considered for small fragment positions or fluorescent dyes with high labeling efficiency. The result is:

[0041] Such as Figure 4 As shown, the primers are divided into five groups: the first group, DYS392, DYS389I, DYS447, DYS389II, DYS438, DYS527a, DYS527b, DYS522, the fluorescent dye marker is FAM; the second group, DYS391, DYS456, DYS19, DYS622, DYS587, DYS385a, DYS385b, the fluorescent dye marker is HEX; the third group, DYS481, DYS437, DYS390, DYS460, DYS643, the fluorescent dye marker is TAMRA; the fourth g...

Embodiment 3

[0042] Example 3 Design of specific primers and adjustment of different primer concentrations

[0043] (1) Determination of the primer sequence of the Y-STR locus

[0044] First, use the UCSC or NCBI website to download the locus sequence through the locus name or the Y chromosome location; secondly, design primers based on the sequences on both sides of the repeat unit of each locus.

[0045] The following points should be paid attention to when designing primers: a. Avoid dimer and hairpin structures between primers, so as not to affect the amplification efficiency of primers; b. Make sure that the primer sequence does not contain SNP sites, especially the 3' end sequence of the primer Stability; c. Primer sequences need to be blasted using NCBI to ensure sequence specificity; d. Ensure that all primers have similar annealing temperatures and have similar amplification efficiencies at the same annealing temperature.

[0046] With the increase of the number of primers in the...

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Abstract

The invention discloses a multi-color fluorescence multiplex amplification kit for amplifying STRgene locuses of a human Y chromosome and application of the kit. Thirty STR gene locuses can be detected in a single amplification system. Compared with the prior art, the kit has the advantages that (1) the gene locuses included in the kit can guarantee the identity of the same family parting and the difference of different family partings due to the advantages high polymorphism, low mutation rate, reasonable frequency distribution and high individual identification; and (2) the kit is extremely high in detection material applicability and can detect various samples.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to molecular genetics, in particular to a multi-color fluorescence composite amplification kit for amplifying the STR locus of human Y chromosome and its application in forensic identification, paternity identification and family investigation. Background technique [0002] Since the 1990s, DNA-STR typing technology has become one of the main means of forensic evidence testing due to its advantages of good specificity, high sensitivity, simple and fast typing, and easy standardization. The short tandem repeat sites (Y-STR) on the Y chromosome commonly used in forensic science are mainly non-recombination regions, so these Y-STRs are completely linked, and all STR loci on the entire Y-STR haplotype can be genetic markers as a whole. Therefore, the use of Y-STR to calculate the probability of individual identification and paternity identification cannot refer to the cumulative method of au...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6876C12Q2600/156
Inventor 吴微微郝宏蕾吕德坚刘冰陈德良王怀锋任文彦苏艳佳梅兴林胡琦郭育林周翔郑卫国
Owner AGCU SCIENTECH
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