Method for identifying gene polymorphism rs2274976 of human MTHFR

Abstract

The invention provides a method for identifying gene polymorphism rs2274976 of human MTHFR. The method comprises the following steps: providing human genome DNA to be tested; using forward primer and reverse primer of the sequence near the gene polymorphism rs2274976 site of the amplified human MTHFR and uses the human genome DNA to be tested as template to perform PCR amplification reaction and obtain an amplification product; using restriction enzyme to perform enzyme cutting to the amplification product; and performing electrophoresis to the enzyme cutting product to identify the gene polymorphism of human MTHFR, wherein the penultimate basic group on the end of the forward primer 3' is a mismatching basic group C to form CCGG or ACCGGT structure with polymorphic G allele in the amplification product, or to form CCAGT structure with polymorphic A allele so that the corresponding incision enzyme can be used for enzyme cleavage. The method of the invention has simple operation, low cost and wide application range.

Description

technical field The present invention relates to methods of identifying single nucleotide polymorphisms. More specifically, the present invention relates to a method for identifying the human MTHFR gene polymorphism rs2274976. Background technique SNP (Single Nucleotide Polymorphism) refers to the DNA sequence variation caused by the change of a single nucleotide, including single base substitution, insertion and deletion. SNP has been widely used in genetic linkage analysis and association analysis of simple and complex diseases, as well as the location of disease susceptibility genes, and guides the cloning of susceptibility genes. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology is a classic method for detecting SNP genotype quickly, easily, accurately and at low cost. The principle of this method is: restriction endonuclease is a class of enzymes that recognize specific DNA sites (usually 4-6 bp) and cut at specific sites. The s...

AI Technical Summary

Problems solved by technology

At present, the endonuclease BsrBI is often used to identify the human MTHFR gene polymorphism rs2274976. The endonuclease is expensive (refer to the following table 1 for the reference price of some endonucleases), which affects the cost of the experiment and is difficult to test in the laboratory. popular use in

And due to the inherent defects of the traditional PCR-RFLP method, it is usually difficult for those skilled in the art to select other restriction endonucleases to identify the human MTHFR gene polymorphism rs2274976

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