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Bisulfite Sequencing for DNA Methylation Analysis of Primary Muscle Stem Cells

  • Kohei Miyata
  • Masashi Naito
  • Tomoko Miyata
  • Sho Mokuda
  • Hiroshi AsaharaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1668)

Abstract

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.

Key words

Epigenetics DNA methylation Methylcytosine Bisulfite conversion Bisulfite sequencing analysis 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Kohei Miyata
    • 1
  • Masashi Naito
    • 2
  • Tomoko Miyata
    • 1
  • Sho Mokuda
    • 1
  • Hiroshi Asahara
    • 1
    • 2
    Email author
  1. 1.Department of Molecular and Experimental MedicineThe Scripps Research InstituteLa JollaUSA
  2. 2.Department of Systems Bio MedicineTokyo Medical and Dental UniversityTokyoJapan

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