Multiplex amplification system based on rapid mutation Y-STR gene loci, method and application

A compound amplification system and gene locus technology, applied in the field of compound amplification system, can solve the problems of increasing cost and time for a single locus, lack of gene frequency and mutation rate of Y-STR loci, etc.

Active Publication Date: 2016-06-08
HUAZHONG UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, these 13 newly discovered high mutation rate Y-STR loci have not been put into practical application due to the lack of gene frequency and mutation rate data of various populations.
And the amplification and detection of a single locus will greatly increase the cost and time required for testing

Method used

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  • Multiplex amplification system based on rapid mutation Y-STR gene loci, method and application
  • Multiplex amplification system based on rapid mutation Y-STR gene loci, method and application
  • Multiplex amplification system based on rapid mutation Y-STR gene loci, method and application

Examples

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Embodiment 1

[0162] Example 1: The primer sequences and concentrations used in the fluorescent multiplex amplification system of Example 1 of the present application are specifically shown in Table 1 below:

[0163] Table 1

[0164]

[0165]

[0166] Example 1 of the present application The method for using the compound amplification system based on the rapid mutation Y-STR locus comprises the following steps:

[0167] 1. Synthesize primers according to the primer sequences in Table 1 above, and label 6-FAM, VIC, NED or PET fluorescent dyes on one primer respectively. Dissolve the primer powder with 1ml of deionized water, and prepare the upstream and downstream primers into a 5μM working solution;

[0168] 2. Mix each pair of primers according to the ratio of concentrations stated in Table 1 to form a primer mixture;

[0169] 3. Genomic DNA was extracted by Chelex-100 method;

[0170] 4. Perform DNA amplification on the ABI2720 amplification instrument;

[0171] 5. Use ABI3130 g...

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Abstract

The invention provides a multiplex amplification system based on rapid mutation Y-STR gene loci, a method and application. The multiplex amplification system comprises 13 gene loci, namely, DYF387S1, DYF399S1, DYF403S1a / b, DYF404S1, DYS449, DYS518, DYS526b, DYS547, DYS570, DYS576, DYS612, DYS626 and DYS627, wherein sequences of amplification primer pairs are shown as SEQ ID NO.1-26, and the gene loci are marked through four fluorescein of 6-FAM, VIC, NED and PET respectively. According to the multiplex amplification system based on the rapid mutation Y-STR gene loci, the method and the application, for the 13 rapid mutation Y-STR gene loci, the primers are designed on the basis of group frequency survey, and the gene loci are integrated into the multiplex reaction system according to specific combination and primer concentration, so that amplification and detection of the gene loci are achieved simultaneously, and the amplification effect is better; when the multiplex amplification system is applied to the detection process, convenience and rapidness are better achieved, the typing result is clear, the correctness, sensitivity and stability of gene locus detection can be guaranteed, and higher individual recognition rate is achieved.

Description

technical field [0001] The invention relates to the detection of polymorphic genetic markers in the human genome, in particular to a compound amplification system based on a rapidly mutated Y-STR gene locus and its application. Background technique [0002] The Y-STR locus is a type of short tandem repeat sequence on the Y chromosome unique to males. The inheritance method is paternal inheritance. It is often used in forensic medicine to identify paternal kinship, investigate sexual crimes, and infer the source of biological samples from the field. aspect. [0003] Now there are about 20 Y-STR loci widely used in the identification of forensic cases in the world, and they are put into use in the form of multiplexed commercial kits, such as the 17-site Yfiler kit from ThermoFisher. In order to ensure the feasibility of parentage identification, the mutation rate of these loci is very low, so as to ensure that the kinship identification will not make false exclusion conclusio...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156C12Q2600/16
Inventor 黄代新张文琼肖超魏巍
Owner HUAZHONG UNIV OF SCI & TECH
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