Bisulfite Sequencing for DNA Methylation Analysis of Primary Muscle Stem Cells

Methods Mol Biol. 2017;1668:3-13. doi: 10.1007/978-1-4939-7283-8_1.


In skeletal muscle, DNA methylation contributes to the suppression of gene expression in several biological processes and diseases. A protocol for the detection of methylated cytosine was thus established based on methylation-sensitive enzymes, immunoprecipitation, and bisulfite conversion. DNA methylation analysis, with bisulfite conversion and sequencing, enables the quantification of methylation at each single base position. Here, we describe a basic method of bisulfite sequencing that can be used to analyze local DNA methylation status to confirm genome-wide DNA methylation analysis or correlation of gene expression regulatory mechanisms.

Keywords: Bisulfite conversion; Bisulfite sequencing analysis; DNA methylation; Epigenetics; Methylcytosine.

MeSH terms

  • Cytosine / metabolism
  • DNA / genetics
  • DNA Methylation / genetics*
  • DNA, Single-Stranded / genetics
  • Genome, Human
  • Genome-Wide Association Study
  • Humans
  • Primary Cell Culture
  • RNA / genetics
  • Satellite Cells, Skeletal Muscle / metabolism*
  • Sequence Analysis, DNA / methods*
  • Sulfites / chemistry*
  • Transcriptome


  • DNA, Single-Stranded
  • Sulfites
  • RNA
  • Cytosine
  • DNA
  • hydrogen sulfite