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Primer set for identifying human Y chromosome typing, and applications, and product using primer set

A Y chromosome and primer set technology, which is applied in biochemical equipment and methods, recombinant DNA technology, and microbial determination/inspection, etc., can solve the problem that the number of Y chromosome STR loci cannot be effectively distinguished from male individuals of the same paternal line, and reduce the Limitations, improve detection efficiency, and reduce detection costs

Inactive Publication Date: 2018-07-27
HANGZHOU JILUO BIOLOGICAL PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The first object of the present invention is to provide a primer set for identifying human Y chromosome typing, the second object of the present invention is to provide the application of the above primer set in identifying human Y chromosome typing, and the third object of the present invention To provide the application of the above-mentioned primer set in the preparation of products for identifying human Y chromosome typing, the fourth object of the present invention is to provide a kit for identifying human Y chromosome typing, so as to alleviate the problems existing in the prior art. Limitations in the number of chromosomal STR loci and technical problems of inability to effectively distinguish between male individuals of the same paternal line

Method used

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  • Primer set for identifying human Y chromosome typing, and applications, and product using primer set
  • Primer set for identifying human Y chromosome typing, and applications, and product using primer set
  • Primer set for identifying human Y chromosome typing, and applications, and product using primer set

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0119] Embodiment 1: the design of the composite amplification primer of 29 STR loci of human Y chromosome

[0120] According to the requirements of the invention, 27 pairs of specific primers were designed to simultaneously amplify 29 Y chromosome STR loci, including: DYS456, DYS576, DYS570, DYS481, DYF387S1, DYS627, DYS549, DYS458, DYS460, DYS437, DYS439, DYS392 , DYS385a, DYS385b, DYS643, DYS393, DYS391, DYS390, DYS635, DYS449, DYS533, DYS438, DYS389I, DYS448, DYS389II, DYS19, GATA, H4 and DYS518.

[0121] The designed primers were determined by Primer3 software, so that the annealing temperature of each locus was as close as possible. In order to avoid possible interactions between primers, AutoDimer v1.0 software was used to detect primer dimers, and detection of no dimers was available. In addition, in order to detect the specificity of the amplified product, BLAST software was used to inquire whether there were amplified sequences other than the target sequence. The d...

Embodiment 2

[0128] Embodiment 2 establishes compound PCR reaction system

[0129]27 pairs of specific primers according to concentration: primer pair 1: 1.2nmol / μl, primer pair 2: 1.2nmol / μl, primer pair 3: 6nmol / μl, primer pair 4: 4nmol / μl, primer pair 5: 6nmol / μl, Primer pair 6: 5nmol / μl, primer pair 7: 2nmol / μl, primer pair 8: 5nmol / μl, primer pair 9: 0.5nmol / μl, primer pair 10: 1nmol / μl, primer pair 11: 6nmol / μl, primer Pair 12: 2.5 nmol / μl, Pair 13: 1.5 nmol / μl, Pair 14: 1.2 nmol / μl, Pair 15: 0.5 nmol / μl, Pair 16: 6 nmol / μl, Pair 17: 1.5 nmol / μl μl, primer pair 18: 2.5 nmol / μl, primer pair 19: 1 nmol / μl, primer pair 20: 0.5 nmol / μl, primer pair 21: 1 nmol / μl, primer pair 22: 1.2 nmol / μl, primer pair 23: 1.2 nmol / μl, primer pair 24: 6 nmol / μl, primer pair 25: 4 nmol / μl, primer pair 26: 6 nmol / μl, primer pair 27: 5 nmol / μl. The above-mentioned primer pairs were configured as primer premix STRtyper-29Y Primer Mix (Plus).

[0130] The reaction system configuration is shown in Table 1:...

Embodiment 3

[0134] Example 3 Application of Human Y Chromosome Typing

[0135] The method for using the kit for identifying human Y chromosome typing comprises the following steps:

[0136] 1. Collect blood spots from father, son and son's uncle in the same family.

[0137] 2. Genomic DNA was extracted by Chelex-100 method, referring to "GA / T 383-2002 Forensic Science DNA Laboratory Test Specification".

[0138] 3. Prepare the reaction system according to Table 2:

[0139] Table 2 PCR reaction system

[0140]

[0141] 4. DNA amplification was carried out on ABI 9700 amplification instrument. The amplification program was: 95°C pre-denaturation for 5 minutes; 94°C denaturation for 10 seconds, 61°C annealing for 1 minute, 30 cycles; 60°C extension for 15 minutes.

[0142] 5. The loading mixture is composed of deionized formamide and molecular weight internal standard (SIZE-525Plus) in the system. Mix 12.5 μL of the loading mixture with 1 μL of the amplification product or STR typer-2...

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Abstract

The invention provides a primer set for identifying human Y chromosome typing, and applications, and a product using the primer set, and relates to the field of biotechnology. According to the presentinvention, the primer set comprises 27 primer pairs and can amplify 29 human Y chromosome STR gene loci, wherein the 29 human Y chromosome STR gene loci contain the multiple rapid mutation sites on the Y chromosome, such that the limitation that the male individuals from the same male line cannot be effectively distinguished is reduced; and the kit for identifying human Y chromosome typing comprises the primer set, can quickly and accurately perform typing identification on human Y chromosome, and has high sensitivity and high specificity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer set for identifying human Y chromosome typing, an application thereof and a product using the same. Background technique [0002] Among the 46 human chromosomes, the Y chromosome is the smallest G-group proximal centromere chromosome, containing about 59,000 kb of DNA. The length of the human Y chromosome is almost only 1 / 3 of the X chromosome. In different races and ethnic groups There is great variability in its length. The entire Y chromosome is mainly composed of the PAR region (pseudoautosomal region) and the male-specific region (male-specific region, MSY). During male meiosis, the pseudoautochromatic region can recombine with the X chromosome. The remaining 95% are specific regions of the Y chromosome. [0003] Gene polymorphism (gene polymorphism) refers to the population in random mating, the same gene locus can have more than two genotypes. DNA site polymorphis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 范嘉庚李洲李鹏飞李璐鲁挺
Owner HANGZHOU JILUO BIOLOGICAL PHARMA CO LTD
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