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Primer for detecting polymorphism of ADH1B gene and ADH2B gene, and application of primer

A gene polymorphism, polymorphic site technology, applied in DNA/RNA fragments, recombinant DNA technology, microbial determination/inspection, etc., can solve the problems of complex operation, high cost, easy pollution, etc., and achieve the detection results. Accurate, simple detection, easy-to-interpret effects

Inactive Publication Date: 2017-06-13
HANGZHOU D A GENETIC ENG
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved in the present invention is to provide a detection primer probe for simultaneously detecting the polymorphism of the ADH1B gene and the ALDH2 gene, so as to overcome the problems of complex operation, easy pollution, and high cost of detecting the polymorphism of the ADH1B gene and the ALDH2 gene in the prior art

Method used

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  • Primer for detecting polymorphism of ADH1B gene and ADH2B gene, and application of primer
  • Primer for detecting polymorphism of ADH1B gene and ADH2B gene, and application of primer
  • Primer for detecting polymorphism of ADH1B gene and ADH2B gene, and application of primer

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Experimental program
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Effect test

Embodiment

[0050] (1) Extraction of DNA from sample to be tested:

[0051] Take 200 μL of peripheral blood and extract DNA according to the instructions of the whole blood genomic DNA extraction kit. The DNA is extracted for subsequent experiments or stored at -20°C.

[0052] (2) Fluorescence quantitative PCR amplification and detection:

[0053] In a 96-well plate, prepare the following reaction system: 12.5μL 2×PCR reaction mixture, 2.5μL primer probe mixture, 50ng template DNA, add water to make up to a total volume of 25μL, and set up a positive control and a negative control at the same time. The amplification procedure in the real-time fluorescent quantitative PCR reaction is: 95°C, 2min; 95°C, 5s, 60°C, 45s, collect fluorescence, 40 cycles.

[0054] (3) Interpretation of results:

[0055] After the reaction is over, use the instrument's own software for data analysis. According to the amplification results, the genotype of the specimen is judged, and the specific test results are as follo...

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Abstract

The invention discloses a primer for detecting polymorphism of an ADH1B gene and an ADH2B gene by a TaqMan-MGB probe method, and application of the primer, and belongs to the technical field of biological detection. Primers and probes as shown in SEQ ID NO:1 to NO:8 are provided. Compared with a sequencing method, the invention has the advantages that PCR products do not need to be subjected to a series of complex subsequent treatment, PCR amplification and detection are conducted synchronously, the whole detection process does not need cover-opening operation, the pollution risk caused by the PCR products is reduced, operation is simple and rapid, the detection time is greatly shortened and the detection cost is reduced. The primer can be used for clinically detecting the polymorphism of the ADH1B gene and the polymorphism of the ADH2B gene, and the detection results can be used for guiding clinical reasonable medication of nitroglycerin and healthy drinking.

Description

Technical field [0001] The invention belongs to the technical field of biological detection, and specifically relates to a method and application for simultaneous detection of ADH1B gene and ALDH2 gene polymorphisms by a TaqMan-MGB probe method. Background technique [0002] Alcohol metabolism-related genes are one of the hot spots in molecular evolution and population genetics. The chemical name of alcohol is ethanol. After drinking, ethanol is absorbed into the blood through the stomach and intestines. 90%-95% of ethanol entering the body is metabolized by the liver, and the rest is excreted with urine, sweat and breathing. Ethanol undergoes two main metabolic reactions in the liver: oxidation from ethanol to acetaldehyde; and oxidation from acetaldehyde to acetic acid. These two metabolic reactions are carried out under the catalysis of enzymes. The enzyme of the former is called alcohol dehydrogenase (ADH) and the enzyme of the latter is called acetaldehyde dehydrogenase (A...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/6858C12Q1/6886C12Q2600/106C12Q2600/156C12Q2531/113C12Q2561/101C12Q2545/113C12Q2563/107
Inventor 任绪义罗英张锋周韵
Owner HANGZHOU D A GENETIC ENG
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