Reagent kit and method for detecting DNA (deoxyribonucleic acid) methylation and application

Abstract

The invention discloses a reagent kit for detecting DNA (deoxyribonucleic acid) methylation. The reagent kit at least comprises biotin-labeled methylated lambda-DNA and streptomycin-enveloped magneticbeads. The invention further discloses application of the reagent kit to detecting methylation of a few DNA samples and a method for detecting the DNA methylation and application of the method. The low sample amount can reach 1 ng. According to the technical scheme, the reagent kit, the application and the method have the advantages that the lowermost initial sample amount required for detectingthe DNA methylation can be obviously lowered, specific reagent kits and experimental equipment can be omitted, accordingly, the cost can be saved, and the reagent kit and the method are high in universality; exogenous DNA pollution can be prevented when methylated DNA is enriched, influence of exogenous DNA information can be prevented if processes for detecting the DNA methylation by means of library construction and high-throughput sequencing are adopted, accordingly, the reagent kit and the method are favorable for further reducing the sequencing cost, and the quality of data can be improved.

Description

technical field [0001] The present invention relates to the technical field of molecular biology, in particular to the field of DNA methylation detection, and more specifically, to a kit, method and application for DNA methylation detection. Background technique [0002] DNA methylation is an important epigenetic modification, which mainly occurs in the CpG island of DNA, and participates in important biological processes such as gene expression regulation, gene imprinting, transposon silencing, X chromosome inactivation, and cancer occurrence. The detection of DNA methylation is widely used in stem cell research, disease diagnosis and treatment and other fields. DNA methylation detection methods can be mainly divided into three categories according to the principles: bisulfite conversion, methylation-sensitive restriction enzyme digestion method, and antibody-enriched methylation site method. Traditional DNA methylation detection methods require microgram-level initial sam...

AI Technical Summary

Problems solved by technology

This method reduces the cost compared to the WGBS method, but the disadvantage is that it can only detect methylation-sensitive restriction enzyme recognition sites, and the restriction enzyme recognition sites are limited, so it cannot be used for genome-wide analysis. Methylation research, and the addition of exogenous DNA can easily affect the sequencing results

Failure to remove additional exogenous DNA may affect sequencing results

[0006] The disadvantages of the existing three technologies are: 1. The cost of sequencing is high, and it is only suitable for analysis of a small number of samples

2. Although the cost is reduced, it cannot be used for genome-wide methylation research, and the exogenous DNA in the system may affect the sequencing results

3. It can detect the methylation level of the whole genome, but it needs to use kits and automatic sample processing system for experiments, which is not universally applicable

Adding exogenous DNA can easily affect the sequencing results

[0007] In addition, the minimum sample input amount required by the above three DNA methylation detection technologies is 20-50ng DNA, which is still not suitable for methylation analysis of a small number of source samples

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