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Method and kit for mutation detection of mitochondrion tRNA<Leu(UUR)>3253T>C

A technology of mitochondria and kits, applied in the field of biomedical engineering, can solve the problems of time-consuming, difficult operation, high cost, etc., and achieve the effect of clear analysis of test results

Active Publication Date: 2016-01-27
WENZHOU MEDICAL UNIV
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Problems solved by technology

[0006] The present invention carries out mitochondrial tRNA by enzyme cutting method Leu(UUR) 3253T>C mutation detection overcomes the shortcomings of existing methods such as time-consuming, difficult operation, and high cost, and provides mitochondrial tRNA Leu(UUR) 3253T>C mutation detection kit, and the above method or the above kit in the detection

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  • Method and kit for mutation detection of mitochondrion tRNA&lt;Leu(UUR)&gt;3253T&gt;C
  • Method and kit for mutation detection of mitochondrion tRNA&lt;Leu(UUR)&gt;3253T&gt;C
  • Method and kit for mutation detection of mitochondrion tRNA&lt;Leu(UUR)&gt;3253T&gt;C

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Embodiment Construction

[0020] 1. DNA extraction: Use the phenol-chloroform-isoamyl alcohol method to extract DNA from blood samples, hair with hair follicles, oral mucosal scrapings or saliva samples.

[0021] 2. Design of specific primers and PCR amplification of specific bands: use Primer5.0 software to design upstream primer 3253F and downstream primer 3253R according to the human mitochondrial gene sequence (NC_012920.1) before and after 3253; Sexual bands were amplified.

[0022] 3. Specific enzyme digestion and electrophoresis identification: the specific restriction endonuclease BstEII is used to digest the above specific bands, and the digested products are subjected to agarose gel electrophoresis to identify whether 3253T>C occurs mutation.

[0023] Specific steps are as follows:

[0024] 1. Collect specimens: detect mitochondrial tRNA as above Leu(UUR) In the gene T3253C mutation method, the whole genome DNA can be quickly extracted from cells, blood and tissue samples. According to the...

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Abstract

The invention discloses a method and a kit for mutation detection of a mitochondrion tRNA<Leu(UUR)>3253T>C. The method includes DNA (deoxyribonucleic acid) extraction, specific primer design, specific band PCR (polymerase chain reaction) amplification, specific restriction enzyme BstEII digestion, agarose gel electrophoresis and the like. Results can be obtained within a few hours, and one enzyme is only capable of identifying one specific deoxyribonucleotide sequence, so that accuracy of the results is realized. The invention further provides a kit for mutation detection of the mitochondrion tRNA<Leu(UUR)>3253T>C related to primary hypertension. The method and the kit for mutation detection of the mitochondrion tRNA<Leu(UUR)>3253T>C have the advantages of low requirements on instruments for detection, low cost of required reagents, simplicity and easiness in operation, stability of the method, high specificity and the like.

Description

technical field [0001] The invention relates to the field of biomedical engineering, in particular to a method for detecting mitochondrial tRNA Leu(UUR) 3253T>C mutation kit and its detection method. Background technique [0002] Cardiovascular disease is the leading cause of death in the world. According to the statistics of the World Health Organization, about 17.3 million people died of cardiovascular disease in 2008. By 2030, almost 23.3 million people will die of cardiovascular disease. Hypertension is number one risk factor for cardiovascular disease. High blood pressure affects more than one-third of adults worldwide, or about 1 billion people, and causes more than 9 million deaths worldwide each year, half of whom die from heart disease and stroke. High blood pressure can also cause kidney failure, blindness, Broken blood vessels and brain damage. Hypertension can be divided into primary hypertension and secondary hypertension according to the cause of the dise...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/683C12Q2521/301C12Q2545/113
Inventor 管敏鑫薛凌郑斌娇唐霄雯林枝
Owner WENZHOU MEDICAL UNIV
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