Multi-component chromatography single nucleotide polymorphism detection method for biologically coding based on general PCR amplification

Abstract

The invention discloses a multi-component chromatography single nucleotide polymorphism detection method for biologically coding based on general PCR (Polymerase Chain Reaction) amplification. The multi-component chromatography single nucleotide polymorphism detection method comprises an allele specific ligase chain reaction, a general PCR amplification reaction, a restriction enzyme digestion reaction, and multi-component quantitative detection for a target gene specific biological coding length fluorescent marker nucleic acid fragment by using liquid chromatography. According to the detection method disclosed by the invention, a connecting product matched with the type of a target gene is generated by using an allele specific ligase chain reaction; a fluorescent marker amplified product of the connecting product is generated by the general PCR amplification reaction; the amplified product is converted into the target gene specific length coding fluorescent marker nucleic acid fragment by using the restriction enzyme digestion reaction; and the multi-component quantitative detection is carried out by the liquid chromatography. The method is less in sample consumption, simple and convenient in operation and lower in cost; and in addition, the method can be used for simultaneously determining multiple components, so that a general technical platform can be provided for biological research on mass screening and antepartum biological research.

Description

technical field [0001] The invention belongs to a biosensing method for detecting single nucleotide polymorphism, which comprises allele-specific ligase chain reaction, general PCR amplification reaction, restriction endonuclease digestion reaction and the detection of target gene by liquid chromatography Multicomponent Quantitative Detection of Type-Specific Bioencoding-Length Fluorescently Labeled Nucleic Acid Fragments. Background technique [0002] The detection of gene polymorphism plays a very important role in the research of molecular biology, pathogen detection, clinical testing, new drug development and human origin and evolution. Many human genetic diseases such as α and β thalassemia are caused by single gene mutations. These genetic diseases are one of the most important birth defects in my country, seriously affecting the population quality of our country, and bringing heavy economic burdens to society and families. Therefore, it is necessary to establish a ra...

AI Technical Summary

Problems solved by technology

However, most of these detection methods are based on nucleic acid molecular hybridization, gel electrophoresis analysis, and multiple labeling of nucleic acid chains. They either have low sensitivity, poor reproducibility, and poor reliability, or low analysis throughput, high cost, and require precision. Advanced instruments cannot meet the needs of economical, accurate and rapid detection