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Non-syndromic hearing loss related mitochondria tRNA <Ser(UCN)> 7444G>A mutation detection method and application thereof

A technology for mutation detection and detection method, applied in the field of genetic engineering, can solve the problems of long time, expensive equipment, complicated operation, etc., and achieve the effect of easy mastery and simple operation.

Inactive Publication Date: 2016-06-01
WENZHOU MAITUO BIOLOGICAL TECH
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Problems solved by technology

However, when using this structure for detection, the operation is complicated, time-consuming, and the equipment is expensive

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  • Non-syndromic hearing loss related mitochondria tRNA &lt;Ser(UCN)&gt; 7444G&gt;A mutation detection method and application thereof
  • Non-syndromic hearing loss related mitochondria tRNA &lt;Ser(UCN)&gt; 7444G&gt;A mutation detection method and application thereof
  • Non-syndromic hearing loss related mitochondria tRNA &lt;Ser(UCN)&gt; 7444G&gt;A mutation detection method and application thereof

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[0031] The embodiments of the present invention will be described in detail below in conjunction with the embodiments, so as to fully understand and implement the process of how to apply technical means to solve technical problems and achieve technical effects in the present invention.

[0032] It should be noted that, in the case of no conflict, the implementations in the present application and the features in the implementations can be combined with each other.

[0033] In one embodiment of the present invention, a kind of mitochondrial tRNA is provided Ser(UCN) Gene G7444A mutation detection kit, the kit includes: reagents for extracting whole-genome DNA from samples, PCR amplification reaction reagents, enzyme digestion reagents, positive standards, negative standards; reagents for extracting whole-genome DNA from samples include: lysate, Solution Ⅰ, phenol-chloroform-isoamyl alcohol mixture and solution Ⅱ; PCR amplification reaction reagents include: dNTP, 10×PCR buffer,...

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Abstract

The invention a mitochondria tRNA <Ser(UCN)> gene 7444A mutation detection method. The method comprises the following steps: applying the phenol-chloroform-isoamyl alcohol method to extract the DNA in a blood specimen, hairs with hair follicles, mouth mucosa doctor-bars or a saliva sample, and designing a forward primer Ser(UCN)F and a reverse primer Ser(UCN)R in front of and behind 7444; expanding a specific strip including 7444; then adopting XbaI for digestion of the specific strip, and performing polyacrylamide gel electrophoresis on a product obtained after digestion to identify whether 7444G>A mutation occurs or not. The method realizes result acquisition within several hours, ensures that one enzyme can only recognize a specific deoxynucleotide sequence, is accurate in result, facilitates building a perfect screening technology for non-syndromic hearing loss related mitochondria tRNA <Ser(UCN)> 7444G>A mutation, and can be popularized in clinical detection.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a mitochondrial tRNA related to non-syndromic deafness Ser(UCN) 7444G>A mutation detection method and kit application. Background technique [0002] Deafness is one of the most common disabilities. There are about 20.04 million deaf people in my country, and the incidence of newborn deafness is 0.1% to 0.3%, of which about 50% are hereditary deafness. However, hereditary deafness has a high degree of genetic heterogeneity, and multiple gene mutations are related to it. Commonly used detection techniques such as restriction endonuclease analysis and gene sequencing are difficult to meet the needs of screening and diagnosis. Many deaf patients are caused by genetic defects, or by increased susceptibility to deafness-causing environmental factors due to genetic defects and polymorphisms. Therefore, research on the molecular etiology of hereditary deafness is very im...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858
Inventor 管敏鑫薛凌郑斌娇唐霄雯范文露
Owner WENZHOU MAITUO BIOLOGICAL TECH
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