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Kit for SNP genotyping of Chinese children short stature susceptibilty genes and utilization method of kit

A susceptibility gene, dwarfism technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of cumbersome operation, long time, limitation and so on

Inactive Publication Date: 2016-02-24
LIUZHOU CITY HEALTHCARE HOSPITAL FOR WOMEN & CHILDREN
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method has disadvantages such as time-consuming, cumbersome operation, and low accuracy.
There are also PCR combined with DNA sequencing methods for the detection of gene polymorphism sites, but this method is limited in the application of large-scale population screening or detection of multiple genes and multiple sites

Method used

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  • Kit for SNP genotyping of Chinese children short stature susceptibilty genes and utilization method of kit
  • Kit for SNP genotyping of Chinese children short stature susceptibilty genes and utilization method of kit
  • Kit for SNP genotyping of Chinese children short stature susceptibilty genes and utilization method of kit

Examples

Experimental program
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Effect test

Embodiment 1

[0082] Example 1: As shown in the accompanying drawings, monitoring of SNP typing of susceptibility genes in patients with dwarfism:

[0083] Sample selection: The samples were derived from the DNA library of dwarf patients in the Children's Health Department of Liuzhou Maternal and Child Health Hospital. Among them, there were 80 males and 80 females. All patients in the bank signed the informed consent before the samples were collected, and promised to cooperate with the follow-up diagnosis and follow-up.

[0084] 1. PCR amplification of 12 segments of 8 dwarfism susceptibility genes with a single pair of primers

[0085] The PCR reaction system is 25 μL, including 10×PCR buffer (Mg 2+ free) 2.5 μL, dNTP mix 0.5 μL (10 mM), 0.8 μL MgCl 2 (50mM), 0.2μL PlatinumTaq DNA polymerase (5U), 0.5μL forward primer per site, 0.5μL reverse primer per site, 1μL DNA template (20mg / L), 19μL ddH 2 O. The PCR reaction conditions were: denaturation at 95°C for 5 min, denaturation at 94°C ...

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Abstract

The invention relates to a kit for SNP genotyping of Chinese children short stature susceptibilty genes and a utilization method of the kit and particularly discloses a simultaneous qualitative and genotyping kit aiming at 12 SNP loci of 8 short stature susceptibilty genes of Chinese children and a utilization method of the simultaneous qualitative and genotyping kit. The method includes: taking a rs17038182 locus of an SPAG17 gene, a rs1378850 locus of a KBTBD gene, a rs1812175 locus of an HHIP gene, a rs10946808 locus of an HIST1H1D gene, a rs8041863 locus of an ACAN gene, a rs6182 locus of a GHR gene, a rs6184 locus of the GHR gene, a rs4410646 locus of the GHR gene, a rs12515480 locus of the GHR gene, a rs2940913 locus of the GHR gene, a rs1976667 locus of an IGF-1R gene and a rs3099 locus of an IHH gene as detection objects, respectively designing amplification primers and extension primers aiming at mutation of each SNP locus, performing multiplex PCR amplification and mark extension aiming at the 12 SNP loci, and performing capillary electrophoresis analysis to obtain genotypes of the 12 SNP loci at the same time. The kit for SNP genotyping has the advantages of convenience in use, simplicity in operation, low cost, high flux, directness and reliability of detection results and applicability to large-scale screening of SNP genotyping of the Chinese children short stature susceptibilty genes.

Description

technical field [0001] The invention relates to the field of SNP typing detection in the technical field of genetic engineering, in particular to a Chinese child dwarfism susceptibility gene SNP typing kit and a use method thereof. Background technique [0002] The incidence rate of shortstature (SS) in children is 3%, which is one of the most common clinical problems for pediatricians, and seriously affects children's future developmental status, psychological state, and future social and job competition. Short stature children's study and future work will be affected to varying degrees. Therefore, in recent years, this problem has attracted increasing attention from society and families at home and abroad, and the number of clinical patients has also increased. Therefore, the exploration of its etiology has special significance. significance. Dwarfism has a complex genetic background. The exploration of susceptibility genes on the chromosome of the disease is still a res...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q2531/113C12Q2535/125C12Q2537/143C12Q2521/319C12Q2521/525
Inventor 严提珍曾婷唐宁曾定元李红辉王麟李伍高李哲涛李静文罗世强
Owner LIUZHOU CITY HEALTHCARE HOSPITAL FOR WOMEN & CHILDREN
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