Electrochemical analysis method of DNA methylation specific sites

Abstract

The invention belongs to electrochemical analysis for anchoring DNA methylation specific sites according to CH3-HS hydrophobic domain liquid phase representation features and discloses an electrochemical analysis method of the DNA methylation specific sites. The electrochemical analysis method has the advantages that the electrochemical method has unique advantages in the researches of DNA methylation due to the fact that the electrochemical method is fast, simple, sensitivity, easy to achieve miniaturization and the like; the physicochemical property that methylated DNA forms a hydrophobic ball using methyl CH3- as the core under a liquid phase condition is utilized to achieve positioning analysis of methylation base sites; the invention provides a series of target sequence (DNA S2, DNA S3, DNA S4, DNA S5, DNA S6, table 1) of known methylation positions.

Description

technical field [0001] The invention belongs to the electrochemical analysis of DNA methylation-specific sites anchored based on the liquid-phase characterization characteristics of CH3-HS hydrophobic domains, and in particular relates to an electrochemical analysis method of DNA methylation-specific sites. Background technique [0002] At present, the existing technology commonly used in the industry is as follows: DNA methylation is the most basic modification method of epigenetics, due to its regulation of gene expression and silencing in cell differentiation, embryonic development, genetic diseases and tumorigenesis, DNA methylation has become the most valuable third-generation genetic marker after restriction fragment polymorphism and DNA point mutation. Studies at home and abroad have successively identified the methylation of genes such as lung cancer CDKN2A, liver cancer ARID2, colon cancer CDKN2A, and leukemia HOXA10 as early markers of tumorigenesis, and found mult...

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Problems solved by technology

[0004] (1) The test results of cloning sequencing are affected by whether the conversion is complete and the sequencing depth. At the same time, the library construction process is complicated, and the acid treatment for a long time may easily lead to the degradation of the target sequence; after the detection, the huge data processing and the special requirements for the detection equipment lead to its Difficult to popularize and apply in clinical laboratory

[0005] (2) MSRE analysis Due to the specificity of restriction endonucleases for base sequence recognition, it can only analyze RE recognition sequences, and the detection range is limited by RE recognition, so this technology can only analyze whether a certain sequence to be tested is In the presence of methylation, the exact site of methylation cannot be accurately determined

[0006] (3) The detection results of the methylation chip technology can only detect whether there is methylation in a known sequence, and cannot clarify the specific site of methylation

At present, Bisulfite-Seq technology, which is applied to genome-wide methylation detection, limits its application in clinical laboratories due to deep sequencing requirements and huge data analysis. MRSE methylation-sensitive restriction endonuclease technology exists The primer design is limited by the recognition sequence of the restriction endonuclease, and the detection result is completely affected by the enzyme digestion

At the same time, MRSE cannot accurately determine the methylation site

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