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Deafness predisposing gene 12S rRNA (ribosomal ribonucleic acid) 1494C>T fluorescence detection kit and application thereof

A deaf susceptibility gene and fluorescence detection technology, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, etc., can solve the problems of many false negatives and false positives, difficult interpretation of results, cumbersome operation, etc., to prevent false positives and false negative, increase Tm value, shorten the effect of primer length

Inactive Publication Date: 2012-07-18
万戈江
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  • Abstract
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Problems solved by technology

These methods have different defects, such as cumbersome operation, difficult interpretation of results, poor repeatability, many false negatives and false positives, etc.

Method used

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  • Deafness predisposing gene 12S rRNA (ribosomal ribonucleic acid) 1494C>T fluorescence detection kit and application thereof
  • Deafness predisposing gene 12S rRNA (ribosomal ribonucleic acid) 1494C>T fluorescence detection kit and application thereof
  • Deafness predisposing gene 12S rRNA (ribosomal ribonucleic acid) 1494C>T fluorescence detection kit and application thereof

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Embodiment 1

[0029] The deaf susceptibility gene 12S rRNA 1494C>T fluorescence detection kit detects mutations and DNA samples from normal individuals, the primers for the detection of the deaf susceptibility gene 12S rRNA 1494C>T, and the primers for the amplification of the Amelogenin locus are all used Labeled with yellow fluorescent dye TAMRA; the internal standard is labeled with red fluorescent dye, and the fluorescent label is ROX. Among them, the primers used for the detection of the deaf susceptibility gene 12S rRNA 1494C>T: primers modified by the incorporation of LNA nucleoside monomers (SEQ NO.2: 5'-GACACACCGCCCGTCTCT-3') and common primers (SEQ NO. .2.0: 5'-GACACACCGCCCGTCTCT-3') for comparison of amplification effects.

[0030] 1. All 100 samples to be tested have been sequenced and detected at the 1494 site of 12S rRNA using the technical method of "DNA extraction-PCR amplification-sequencing", including 2 mutation samples.

[0031] 2. Genomic DNA extraction from samples

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Abstract

The invention discloses a fluorescence detection kit for detecting deafness predisposing gene 12S rRNA (ribosomal ribonucleic acid) 1494C>T. The kit comprises a reagent before amplification and a reagent after amplification, wherein the reagent before amplification comprises a reaction mixture of a PCR (polymerase chain reaction) buffer solution, MgCl2 and dNTPs (deoxyribonucleoside triphosphate), Taq enzyme, ultrapure water, and a primer mixture for high-specific amplification of 12SrRNA1494C>T and amelogenin locus; and the reagent after amplification comprises an allelic ladder and an internal standard. According to the invention, 12SrRNA1494C>T and amelogenin locus are used as detecting objects, so that the fluorescence detection kit has important meaning to detection of deafness predisposing genes, particularly screening of newborn deafness genes.

Description

technical field [0001] The invention relates to a fluorescent detection kit for detecting the susceptibility gene 12S rRNA 1494C>T and the Amelogenin gene locus of deafness, and belongs to the technical field of biological detection. Background technique [0002] Worldwide studies on the pathogenic factors of hearing and speech disabilities show that about 60-80% of the patients are caused by genetic factors, and clinical research data from developed countries show that hereditary deafness accounts for about 80% of deafness patients. Therefore, in the past ten years, the research on the pathogenesis and molecular epidemiology of hereditary deafness has become one of the most important contents of deaf research. With the completion of the Human Genome Project, great progress has been made in the positioning and cloning of deafness genes. The molecular genetics research and molecular epidemiological data of deafness have enabled researchers to gradually realize that mutatio...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 万戈江魏宏泉步讯夏子芳
Owner 万戈江
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