Kit for detecting acetaldehyde dehydrogenase 2 (ALDH2) gene polymorphism by pyro-sequencing method and method

A pyrosequencing method and gene polymorphism technology, applied in the field of molecular biology, can solve the problems of reduced enzyme activity, reduced angina pectoris effect, blocked biotransformation process, etc., and achieves the effect of small sample size and easy operation.

Inactive Publication Date: 2013-01-16
周宏灏
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the ALDH2 gene is mutated at the rs671 site, the enzyme activity decreases, the biotransformation process of nitroglycerin in the body is blocked, and the generation of effective active product NO is reduced, which in turn leads to a decrease in the effect of nitroglycerin in treating angina pectoris.

Method used

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  • Kit for detecting acetaldehyde dehydrogenase 2 (ALDH2) gene polymorphism by pyro-sequencing method and method
  • Kit for detecting acetaldehyde dehydrogenase 2 (ALDH2) gene polymorphism by pyro-sequencing method and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] ALDH2-pyroF (upstream primer): 5′-GATGTGTTTGGAGCCCAGTC-3′ (SEQ ID NO.2);

[0031] ALDH2-pyroR (downstream primer): 5′-CAGGTCCCACACTCACAGTTTTC-3′ (SEQ ID NO.3);

[0032] Sequencing primer: 5′-GCTGCAGGCATACAC-3′ (SEQ ID NO.4);

[0033] 1. DNA extraction

[0034] 1.1 The preparation and inspection of reagent materials before the experiment are as follows:

[0035] (1) Check the shelf life of the kit and ensure that ethanol has been added to Wash Buffer 1 and 2, and tick the corresponding mark on the bottle; (2) Isopropanol (if not available, it can be replaced by absolute ethanol) and 75% ethanol ; (3) 1.5mL Eppendorf tubes and various pipette tips within the validity period of autoclaving.

[0036] 1.2 Take out the EDTA anticoagulant tube containing whole blood from the refrigerator at 4°C, and mix it upside down several times;

[0037] 1.3 Make a mark on the 1.5mL Eppendorf tube corresponding to the unique identification of the specimen;

[0038] 1.4 Pipette 900uL C...

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Abstract

The invention discloses a kit for detecting acetaldehyde dehydrogenase 2 (ALDH2) gene polymorphism by a pyro-sequencing method and a method. By the kit, the ALDH2 gene polymorphism, particularly RS671 single nucleotide polymorphism can be detected. The kit comprises primers which are shown as SEQ ID NO.2-4. By the kit, the accurate, quick and high-throughout detection of ALDH2 gene polymorphism can be realized, so that the alcohol tolerance can be effectively predicted and prevented, and the clinical individual administration of nitroglycerin can be guided.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a kit and a method for detecting ALDH2 gene polymorphism by pyrosequencing. Background technique [0002] Alcohol tolerance (Alcohol Tolerance) mainly refers to the decreased sensitivity of the body to ethanol in alcoholic beverages or food, including direct tolerance and induced tolerance. Direct alcohol tolerance is related to the size of the individual, and induced tolerance refers to the increase of alcohol tolerance caused by long-term drinking. Studies have shown that environmental factors and genetic factors jointly affect individual differences in alcohol consumption. Alcohol dependence and alcohol allergy are largely determined by genetic factors. For example, acetaldehyde dehydrogenase 2 (ALDH2) is the main metabolic enzyme of alcohol oxidative metabolism, alcohol tolerance It is related to the enzyme activity of ALDH2, and its polymorphism is also the main...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 周宏灏
Owner 周宏灏
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