Construction method of targeted methylation sequencing library and kit

Abstract

The invention discloses a construction method of a targeted methylation sequencing library and a kit. The construction method comprises the steps of construction of a methylation library and hybridization capture of a targeted methylation library, and the construction of the methylation library comprises the following steps: carrying out terminal repair on fragment DNA, carrying out methylation linker connection, converting unmethylated C basic groups in linker connection products into U basic groups, carrying out PCR amplification by using amplification enzyme capable of recognizing the U basic groups, and carrying out hybridization capture on the targeted methylation library. A methylation library is obtained; the target methylation library hybridization capture comprises the steps of taking a methylation library and carrying out sample mixing, concentration, methylation library hybridization, methylation library thermal elution, methylation library normal-temperature elution, target methylation library amplification and target methylation library purification, and the purified target methylation library is the target methylation sequencing library. According to the technical scheme, the construction method of the targeted methylation sequencing library is provided, and the method is more suitable for targeted capture of the methylation library.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a method and kit for constructing a targeted methylation sequencing library. Background technique [0002] DNA methylation is related to many physiological and pathological factors and carries a large amount of biological information. Methylation modification in the promoter region inhibits gene transcription and stabilizes epigenetic marks; methylation modification in the gene body increases gene expression and stimulates transcription elongation. Methylation modification of repetitive sequence regions maintains genome stability. Compared with normal cells, there are usually two forms of DNA methylation changes in tumor cells, hypomethylation (hypomethylation) and hypermethylation (hypermethylation), and hypomethylation modification in repetitive regions leads to chromosomal instability. The silenced gene loses methylation modification and is activated, which leads to the ...

AI Technical Summary

Problems solved by technology

Targeted methylation sequencing based on liquid-phase hybridization capture technology is divided into two categories, one is capture first and then transform, the advantage is that the data capture preference is low, and the disadvantage is that the requirement for template input is high, which is not conducive to low concentration Sample library construction; the other is to transform first and then capture, which requires less template. The disadvantage is that the capture has a certain preference, and it has higher requirements for the hybrid capture reaction system and reaction conditions.

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