Deafness susceptibility gene mutation detection kit as well as preparation method and application thereof

A technology for detection kits and genes, which can be used in biochemical equipment and methods, determination/inspection of microorganisms, bioreactor/fermenter combination, etc.

Active Publication Date: 2015-01-07
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a simple and fast, high sensitivity, high reliability and low cost multi-colo

Method used

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  • Deafness susceptibility gene mutation detection kit as well as preparation method and application thereof
  • Deafness susceptibility gene mutation detection kit as well as preparation method and application thereof
  • Deafness susceptibility gene mutation detection kit as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] Example 1: Use this kit to detect reference products of known genotypes HHL W (wild type), HHL M1 (c.35delG homozygous mutant), HHL M2 (c.167delT homozygous mutant), HHL M3 ( c.176_191del16 homozygous mutant), HHL M4 (c.538C>T homozygous mutant), HHL M5 (c.547G>A homozygous mutant), HHL M6 (c.235delC homozygous mutant), HHL M7 (c.299_300delAT homozygous mutant), HHL M8 (m.1555A>G homozygous mutant), HHL M9 (m.1494C>T homozygous mutant), HHL M10 (m.7444G>A homozygous mutant type), HHL M11 (m.7445G>A homozygous mutant), HHL M12 (m.961T>G homozygous mutant), HHL M13 (c.919-2A>G homozygous mutant), HHL M14 ( c.2162C>T homozygous mutant), HHL M15 (c.2168A>G homozygous mutant), HHL M16 (c.1174A>T homozygous mutant), HHL M17 (c.1226G>A homozygous mutant type), HHL M18 (c.1229C>T homozygous mutant), HHL M19 (c.1707+5G>A homozygous mutant), HHL M20 (c.1975G>C homozygous mutant), HHL M21 ( c.749T>C homozygous mutant), HHL M22 (c.754T>C homozygous mutant), HHL M23 (c.2027T>A hom...

Embodiment 2

[0112] Embodiment 2: Utilize this kit to detect 148 cases of DNA samples of known genotype, and set standard control and negative control at the same time, including the following steps:

[0113] 1) Use the gene of the sample to be tested and the standard control gene as templates to prepare PCR reaction solutions: 25 μL of PCR reaction solutions A, B, C, and D respectively include: 5 μL of DNA template (negative control is water), 1 × PCR buffer , 3.0 mM MgCl 2 , 1U Taq DNA polymerase, dATP, dCTP, dGTP, dTTP, dUTP each 0.2mM, and the corresponding primer probe.

[0114] 2) Place the above-mentioned PCR reaction tube containing the PCR reaction solution on a fluorescent PCR instrument (Bio-Rad CFX 96) (for PCR amplification and melting curve analysis, the specific reaction procedure is:

[0115] (1) 50°C for 2 minutes, 95°C for 10 minutes;

[0116] (2) 95°C for 15s→65~56°C for 15s→76°C for 20s, 10 cycles, of which 65~56°C for 15s drops by 1°C per cycle;

[0117] (3) 95°C fo...

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Abstract

The invention discloses a deafness susceptibility gene mutation detection kit as well as a preparation method and application thereof, and relates to gene mutation detection. The kit comprises a kit body, a cover, a partition, an HHL (Hereditary Hearing Loss) PCR (Polymerase Chain Reaction) mixed liquor bottle A, an HHL PCR mixed liquor bottle B, an HHL PCR mixed liquor bottle C, an HHL PCR mixed liquor bottle D, an HHL enzyme mixed liquor bottle, an HHL standard control bottle, and an HHL negative control bottle. The preparation method comprises the following steps: preparing an amplification reagent and a contrast reagent firstly, arranging the amplification reagent and the contrast reagent in the bottles and finally, obtaining the deafness susceptibility gene mutation detection kit. The kit can be applied to screening of hereditary deafness genes and identification of deafness causing genes, so as to prevent deafness.

Description

technical field [0001] The invention relates to gene mutation detection, in particular to a deafness susceptibility gene mutation detection kit and its preparation method and application. Background technique [0002] According to statistics, there are currently more than 4,000 known genetic diseases, including thalassemia, congenital stupidity, and deafness. Among them, deafness is a common disease that seriously affects the quality of human life, and is the most common cause of communication disorders. Among all deafness-causing factors, genetic factors are the main reason for the birth of deaf children, accounting for as high as 50% to 60%, which has high genetic heterogeneity. In addition, among a large number of patients with delayed hearing loss, many patients are also caused by their own genetic defects, or due to increased susceptibility to deafness-causing environmental factors due to genetic defects and polymorphisms. [0003] In recent years, with the implementa...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12M1/00
CPCC12Q1/6883C12Q2600/156
Inventor 黄秋英李庆阁王旭东
Owner XIAMEN UNIV
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