Unit

UNIT 7.24 Direct Bisulfite Sequencing for Examination of DNA Methylation with Gene and Nucleotide Resolution from Brain Tissues

  1. R. Ryley Parrish1,2,
  2. Jeremy J. Day1,2,
  3. Farah D. Lubin1,2

Published Online: 1 JUL 2012

DOI: 10.1002/0471142301.ns0724s60

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Parrish, R. R., Day, J. J. and Lubin, F. D. 2012. Direct Bisulfite Sequencing for Examination of DNA Methylation with Gene and Nucleotide Resolution from Brain Tissues. Current Protocols in Neuroscience. 60:7.24:7.24.1–7.24.12.

Author Information

  1. 1

    Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of Alabama at Birmingham, Birmingham, Alabama

  2. 2

    These authors contributed equally to this work.

Publication History

  1. Published Online: 1 JUL 2012
  2. Published Print: JUL 2012

Abstract

DNA methylation is an epigenetic modification that is essential for the development and mature function of the central nervous system. Due to the relevance of this modification to the transcriptional control of gene expression, it is often necessary to examine changes in DNA methylation patterns with both gene and single-nucleotide resolution. Here, we describe an in-depth basic protocol for direct bisulfite sequencing of DNA isolated from brain tissue, which will permit direct assessment of methylation status at individual genes as well as individual cytosine molecules/nucleotides within a genomic region. This method yields analysis of DNA methylation patterns that is robust, accurate, and reproducible, thereby allowing insights into the role of alterations in DNA methylation in brain tissue. Curr. Protoc. Neurosci. 60:7.24.1-7.24.12. © 2012 by John Wiley & Sons, Inc.

Keywords:

  • bisulfite sequencing;
  • DNA methylation;
  • epigenetics;
  • central nervous system