High-throughput sequencing of methylated cytosine enriched by modification-dependent restriction endonuclease MspJI

BMC Genet. 2013 Jun 18:14:56. doi: 10.1186/1471-2156-14-56.

Abstract

Background: As a well-known epigenomic modification, DNA methylation is found to be common in plants and plays an important role in many biological processes. Relying on the unique feature of methylation-dependent digestion, the family of methylation-requiring restriction-like endonuclease, such as MspJI and its homologs, was suggested for a potential usage in methylation detection.

Results: In this study, we combine MspJI digestion and electrophoretic band selection with next generation high-throughput sequencing technology to detect 5-methylcytosines in Arabidopsis genome. By developing a bioinformatics workflow to attribute the CNNR sites recognized by MspJI to the reference genome, we fulfilled the systematic assessment of this method.

Conclusions: According to the assessment, here we provide the method for generating a detailed map of plant methylome that could be feasible, reliable and economical in methylation investigation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics
  • Cytosine / metabolism*
  • DNA Methylation
  • DNA Restriction Enzymes / metabolism*
  • High-Throughput Nucleotide Sequencing*

Substances

  • Cytosine
  • DNA Restriction Enzymes