Hi-Meth: a platform for high-throughput detection of site-specific DNA methylation.

Huiying LI; Qing Liu; Min Guo; Kejian Wang; Changjie Yan; Chun Wang

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Hi-Meth: a platform for high-throughput detection of site-specific DNA methylation.

Author: Huiying LI ¹ ; Qing LIU ² ; Min GUO ¹ ; Kejian WANG ² ; Changjie YAN ¹ ; Chun WANG ²
Author Information

1. Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding, Jiangsu Key Laboratory of Crop Genetics and Physiology/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural college of Yangzhou University, Yangzhou University, Yangzhou 225009, Jiangsu, China.
2. State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310006, Zhejiang, China.
Publication Type:Journal Article
Keywords: 5-methylcytosine; Hi-Meth; detection of methylation specific site; high-throughput sequencing platform
MeSH: DNA Methylation; Epigenesis, Genetic; Epigenomics; High-Throughput Nucleotide Sequencing/methods*; Polymerase Chain Reaction; Sequence Analysis, DNA/methods*
From: Chinese Journal of Biotechnology 2022;38(8):3049-3061
CountryChina
Language:Chinese
Abstract: Cytosine methylation is one of the major types of DNA epigenetic modifications and plays an important role in maintaining normal cell function and regulating gene expression. Bisulfite sequencing PCR (BSP) based cloning and sequencing is a general method for detecting DNA methylation at specific sites, which can clarify the methylation status of each CpG site in the target fragment. However, this method requires large amounts of single-clonal sequencing, which is complicated to operate, time consuming and expensive. Therefore, the development of an accurate, efficient and convenient DNA methylation detection technology is of great significance to improve the efficiency of epigenetic research. Based on the high-throughput mutation detection platform Hi-TOM (high-throughput tracking of mutations) developed by our group, we further established a site-specific DNA methylation high-throughput detection platform Hi-Meth (High-throughput Detection of DNA Methylation). After bisulfite treatment of DNA samples, the specific site-specific DNA methylation analysis results could be obtained through the Hi-Meth platform by performing only one round of PCR amplification. Using the Hi-Meth platform, the DNA methylation status of two promoter regions of rice were detected. The DNA methylation results from Hi-Meth were consistent with the results from BSP-based method. Thus, site-specific DNA methylation analysis results could be obtained accurately and conveniently through the Hi-Meth platform. In conclusion, Hi-Meth provides an important methylation detection platform for specific DNA regions, which has important significance for epigenetic research.