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Compound amplification kit for InDel genetic polymorphic sites of human euchromosome and Y chromosome and application thereof

A genetic polymorphism and autosomal technology, applied in the direction of recombinant DNA technology, biochemical equipment and methods, DNA / RNA fragments, etc., can solve the problems of low identification efficiency, poor detection effect, and few detection sites

Active Publication Date: 2017-06-20
GUANGZHOU CRIMINAL SCI & TECH RES INST +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But this prior art has the following deficiencies: (1) adopt four-color fluorescence system, identification efficiency is low; (2) partial PCR amplification product is greater than 200bp, for the inspection material of degradation, detection effect is not good; (3) detection Few sites, low degree of crowd recognition

Method used

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  • Compound amplification kit for InDel genetic polymorphic sites of human euchromosome and Y chromosome and application thereof
  • Compound amplification kit for InDel genetic polymorphic sites of human euchromosome and Y chromosome and application thereof
  • Compound amplification kit for InDel genetic polymorphic sites of human euchromosome and Y chromosome and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A human autosome and Y chromosome InDel genetic polymorphic site composite amplification kit, said kit contains 47 pairs of autosome InDel site locus specific amplification primers, 2 pairs of Y chromosome InDel site genes Specific amplification primers for loci and 1 pair of specific amplification primers for sex identification genes; the InDel loci are: InDel-01 (rs3067397), InDel-02 (rs10607699), InDel-03 (rs71852971), InDel-04(rs67487831), InDel-05(rs140683187), InDel-06(rs5787309), InDel-07(rs34287950), InDel-08(rs67100350), InDel-09(rs769299), InDel-10(rs3076465), InDel -11(rs67939200), InDel-12(rs145941537), InDel-13(rs67264216), InDel-14(rs35453727), InDel-15(rs35065898), InDel-16(rs60575667), InDel-17(rs66911131), InDel 18(rs61490765), InDel-19(rs34419736), InDel-20(rs34421865), InDel-21(rs66739142), InDel-22(rs3029189), InDel-23(rs33971783), InDel-24(rs72085595), InDel-2 (rs34529638), InDel-26(rs11277697), InDel-27(rs10558392), InDel-28(rs538690481), InDel-29...

Embodiment 2

[0051] The kit described in Example 1 was used to detect old bone samples.

[0052] Amplification system:

[0053]

[0054]

[0055] Among them, the extraction of DNA samples was carried out with reference to the "GA / T 383-2002 Forensic Science DNA Laboratory Test Specification", and all such samples were extracted by the magnetic bead method.

[0056] Amplification procedure:

[0057]

[0058] Amplified products are detected by fluorescence on a genetic analyzer:

[0059] The sample loading mixture is composed of deionized formamide and the molecular weight internal standard AGCU Marker SIZ-500 in the system [(0.5 μl AGCU Marker SIZ-500 (Wuxi Zhongde Meilian Biotechnology Co., Ltd.)) × (injection number) + (12 μl Deionized formamide) × (injection number)]. Mix 12.5 μl of the loading mixture with 1 μl of the amplification product in this kit, avoiding the generation of air bubbles. Denature at 95°C for 3 minutes, ice-bath for 3 minutes, and perform electrophoresis...

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Abstract

The invention discloses a compound amplification kit for InDel genetic polymorphic sites of human euchromosome and Y chromosome and application thereof. The kit comprises 47 pairs of euchromosome InDel site loca, 2 pairs of Y chromosome InDel site loca and a pair of specific amplification primers of a sex determination gene. The kit can be used for human individual recognition, paternity identification and degraded detection material recognition. The kit comprises 49 InDel sites which are relatively balanced in types and one sex determination gene. By adopting a six-colored STR fluorescence detection system, the kit is higher than the disclosed legal medical InDel detection kit. The kit disclosed by the invention is suitable for detecting Chinese groups. An amplification system of 50 sites is short in amplification fragment, and the amplicon is controlled at 200bp, so that the system is suitable for InDel typing of high degraded detection material; the amplification system improves the detection numbers of sites in the degraded detection material; the two Y-InDel sites introduced into the system plays an auxiliary judging role on the sex determination gene Amelogenin.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to gene detection by using molecular markers, in particular to a composite amplification kit for human autosome and Y chromosome InDel genetic polymorphism sites and its application. Background technique [0002] Paternity testing and individual identification are the main contents of forensic DNA identification. Short tandem repeat (short tandem repeat, STR) typing technique is a commonly used technique at present. The STR locus is a short tandem repeat polymorphic genetic marker with a core sequence of 2-6 bp. In the early 1990s, the STR locus was first used as an important genetic marker in human paternity testing. Since then, forensic evidence identification has entered the era of DNA identification. With the wide application of STR loci in forensic DNA analysis, its defects have also attracted increasing attention. For example, the high mutation rate of STR loci is not conduciv...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 刘长晖陈玲刘超于安路杜蔚安郭育林陈林丽刘宏李越郑卫国
Owner GUANGZHOU CRIMINAL SCI & TECH RES INST
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