Method for typing optimization of SNP sites of respiratory system disease genes

Abstract

The invention discloses a method for typing optimization of SNP sites of respiratory system disease susceptibility genes. The method comprises the following steps: 1) extracting DNA; 2) performing PCRamplification on the DNA; 3) purifying a PCR amplification product; 4) extending the purified PCR amplification product; 5) purifying the extension product; 6) performing loading to a sequencer; and7) analyzing fluorescence labeling and length information of the extension product. Based on an SNaPshot technology, PCR amplification primers and extension primers with different lengths are designedfor different SNP sites to achieve typing of multiple SNPs in one reaction system, and typing can be conducted at 14 sites at the same time. Due to the fact that the method adopts four-color fluorescence labeling, classification can be performed according to various SNP types, and besides, insertion and deletion can be analyzed. After genotyping of a respiratory system disease patient is obtainedthrough analysis, prevention before illness of the user and symptomatic medicine application of a doctor after the disease occurs are facilitated.

Description

technical field [0001] The invention relates to the technical field of gene identification, in particular to a method for typing and optimizing SNP sites of respiratory disease genes. Background technique [0002] Respiratory system diseases are the most common diseases in my country. The two-week prevalence rate, two-week visit rate, and number of hospitalizations of urban and rural residents rank first for a long time, and the resulting deaths rank first to fourth in the cause of death, and the disease burden ranks third. , has become one of the most prominent public health and medical problems in our country. Chronic respiratory disease is one of the "four major chronic diseases" defined by WHO. Lung cancer has become the number one tumor in my country, tuberculosis will become the number one infectious disease in my country, and pneumoconiosis accounts for 90% of occupational diseases. In addition, new outbreaks Public health events such as respiratory infectious diseases...

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Problems solved by technology

However, the above methods have certain disadvantages: SSCP can only use non-denatured glue, and the experimental conditions are harsh, requiring room temperature conditions, which are difficult to achieve in general laboratories; the experimental process of CAPs is cumbersome, and an experimental procedure takes a long time; MassARRAY mass spectrometry The cost of TaqMan probe method and other chip methods is quite high, and it is difficult for ordinary laboratories to afford

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