DNA Methylation Detection: Bisulfite Genomic Sequencing Analysis

Part of the Methods in Molecular Biology book series (MIMB, volume 791)

Abstract

DNA methylation, which most commonly occurs at the C5 position of cytosines within CpG dinucleotides, plays a pivotal role in many biological procedures such as gene expression, embryonic development, cellular proliferation, differentiation, and chromosome stability. Aberrant DNA methylation is often associated with loss of DNA homeostasis and genomic instability leading to the development of human diseases such as cancer. The importance of DNA methylation creates an urgent demand for effective methods with high sensitivity and reliability to explore innovative diagnostic and therapeutic strategies. Bisulfite genomic sequencing developed by Frommer and colleagues was recognized as a revolution in DNA methylation analysis based on conversion of genomic DNA by using sodium bisulfite. Besides various merits of the bisulfite genomic sequencing method such as being highly qualitative and quantitative, it serves as a fundamental principle to many derived methods to better interpret the mystery of DNA methylation. Here, we present a protocol currently frequently used in our laboratory that has proven to yield optimal outcomes. We also discuss the potential technical problems and troubleshooting notes for a variety of applications in this field.

Key words

DNA methylation Epigenetics Bisulfite genomic sequencing 

Notes

Acknowledgments

This work was supported in part by grants from the National Cancer Institute (R01 CA 129415), the Susan G. Komen for the Cure and a Postdoctoral Award (PDA) sponsored by the American Institute for Cancer Research (AICR).

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of BiologyUniversity of Alabama at BirminghamBirminghamUSA

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