DNA methyltransferase accessibility protocol for individual templates by deep sequencing

Methods Enzymol. 2012:513:185-204. doi: 10.1016/B978-0-12-391938-0.00008-2.

Abstract

A single-molecule probe of chromatin structure can uncover dynamic chromatin states and rare epigenetic variants of biological importance that bulk measures of chromatin structure miss. In bisulfite genomic sequencing, each sequenced clone records the methylation status of multiple sites on an individual molecule of DNA. An exogenous DNA methyltransferase can thus be used to image nucleosomes and other protein-DNA complexes. In this chapter, we describe the adaptation of this technique, termed Methylation Accessibility Protocol for individual templates, to modern high-throughput sequencing, which both simplifies the workflow and extends its utility.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Computational Biology / methods*
  • CpG Islands
  • DNA (Cytosine-5-)-Methyltransferases / metabolism*
  • DNA / genetics
  • DNA / metabolism*
  • DNA Methylation*
  • Gene Library
  • Genome, Human
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Neurofibromin 1 / genetics*
  • Neurofibromin 1 / metabolism
  • Nucleosomes / genetics
  • Nucleosomes / metabolism*
  • Polymerase Chain Reaction / methods
  • Promoter Regions, Genetic
  • Sequence Analysis, DNA
  • Sulfites / metabolism
  • Templates, Genetic

Substances

  • Neurofibromin 1
  • Nucleosomes
  • Sulfites
  • DNA
  • DNA (Cytosine-5-)-Methyltransferases
  • hydrogen sulfite