Method for detecting gene type of polymorphic site rs671 of gene ALDH2 (Acetaldehyde Dehydrogenase 2) and kit

A polymorphic site and genotype technology, used in biochemical equipment and methods, recombinant DNA technology, and microbial assay/inspection. site and other problems, to achieve the effect of simple operation, improved accuracy, and low detection cost

Inactive Publication Date: 2017-01-11
GUANGDONG PHARMA UNIV
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AI Technical Summary

Problems solved by technology

In addition, the existing scheme using AcuI enzyme digestion also lacks an internal control enzyme digestion site, and there is a possibility of genotype misjudgment
[0020] As for TspRI (TscAⅠ), its price is the same as that of MboⅡ, but the enzyme inactivation operation after enzyme digestion is relatively cumbersome. The instructions suggest using phenol / chloroform extraction and alcohol precipitation, unlike MboⅡ, which can be inactivated at 65°C for 5 minutes.

Method used

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  • Method for detecting gene type of polymorphic site rs671 of gene ALDH2 (Acetaldehyde Dehydrogenase 2) and kit
  • Method for detecting gene type of polymorphic site rs671 of gene ALDH2 (Acetaldehyde Dehydrogenase 2) and kit
  • Method for detecting gene type of polymorphic site rs671 of gene ALDH2 (Acetaldehyde Dehydrogenase 2) and kit

Examples

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Embodiment 1

[0051] In this example, a pair of primer pairs for detecting the genotype of the rs671 site of the ALDH2 gene was designed for the restriction enzyme MboII, and a detection method for the genotype of the rs671 polymorphic site of the ALDH2 gene was established, as follows:

[0052] 1. Genomic DNA Extraction

[0053] Using the buccal swab genomic DNA extraction kit, the genomic DNA was extracted according to the operation steps in the manual. The DNA source is not limited thereto, and any somatic cell-derived DNA is acceptable.

[0054] 2. Primer Design

[0055] Search the DNA sequence related to the rs671 polymorphic site of the ALDH2 gene in the dbSNP database (refer to http: / / www.ncbi.nlm.nih.gov / snp?term=rs671), and retrieve 1000 bp before and after the rs671 polymorphic site of the ALDH2 gene (refer to http: / / www.ncbi.nlm.nih.gov / nuccore / NG_012250.1), the 42421st base is the polymorphic site base, and this sequence shows the base g of the wild-type allele. The accession...

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Abstract

The invention discloses a method for detecting a gene type of a polymorphic site rs671 of a gene ALDH2 (Acetaldehyde Dehydrogenase 2) and a kit. A specific 'inner quality control' mechanism is introduced in a process of detecting the gene type of the polymorphic site rs671 of the gene ALDH2, i.e. an inner contrast cleavage site is introduced into a PCR product; the situation of incomplete cleavage can be identified according to a cleavage atlas; and furthermore, under a condition of the incomplete cleavage and a condition that partial cleavage bands are blurred, the gene type still can be accurately judged, so that the accuracy of a detection result is greatly improved.

Description

technical field [0001] The invention belongs to the field of molecular detection, and in particular relates to a method and a kit for detecting the genotype of the rs671 polymorphic site of the ALDH2 gene. Background technique [0002] Aldehyde dehydrogenase 2 (ALDH2) is a key enzyme in alcohol metabolism, which can catalyze the dehydrogenation of toxic and carcinogenic acetaldehyde produced by ethanol metabolism into non-toxic acetic acid. In addition, ALDH2 also has nitrate esterase activity, which is the key to the formation of nitric oxide, an effective metabolite of nitroglycerin. [0003] The ALDH2 gene contains 13 exons, and there is a single nucleotide polymorphism (Singlenucleotide polymorphism, SNP) site in exon 12, and its dbSNP number is rs671, which is the 1510th base of the open reading frame. → A mutation (G1510A), resulting in the 487th glutamic acid in the polypeptide chain being changed to lysine (Glu487Lys). If an individual carries the Glu487Lys mutation...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q1/6858C12Q2600/156C12Q2545/101
Inventor 何震宇邹碧如黄丽玲杨泽民谢钰铃梁焯梅曾雪燕
Owner GUANGDONG PHARMA UNIV
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