Cited by 358Epigenetics and DNA MethylationGenetic Syndromes and ImprintingDNA and Nucleic Acid Chemistry

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DNA Methylation Detection: Bisulfite Genomic Sequencing Analysis

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Li, Yuanyuan, Tollefsbol, Trygve O.·2011·Methods in Molecular Biology

DOI 10.1007/978-1-61779-316-5_2PMC PMC3233226

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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.

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Authors: Li, Y., & Tollefsbol, T. O.
Affiliations: University of Alabama at Birmingham (US)
Corresponding author: Tollefsbol, Trygve O.
ORCID: Li, Yuanyuan
Tollefsbol, Trygve O.
Venue: Methods in Molecular Biology
Publisher: Humana Press
Published: 2011
Volume: 791
Pages: 11-21
ISSN: 1064-3745 (ISSN-L)
1940-6029
DOI: 10.1007/978-1-61779-316-5_2
PMC: PMC3233226
OpenAlex: W1522996100
Cited by: 352
References: 26
Citations by year: 2026: 25 · 2025: 34 · 2024: 41 · 2023: 32 · 2022: 35 · 2021: 41 · 2020: 36 · 2019: 23 · 2018: 23 · 2017: 22 · 2016: 16 · 2015: 13 · 2014: 6 · 2013: 3 · 2012: 1
FWCI: 4.57
Type: book chapter
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: www.springernature.com/gp/researchers/text-and-data-mining (tdm)
www.springernature.com/gp/researchers/text-and-data-mining (vor)
Locations: Methods in molecular biology (publishedVersion)
PubMed (publishedVersion)
Europe PMC (PubMed Central) (acceptedVersion)
PubMed Central (acceptedVersion)
Funders: Susan G. Komen
National Cancer Institute (R01CA129415)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic Syndromes and Imprinting
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
Keywords: DNA methylation, bisulfite sequencing, CpG, genomic DNA, aberrant methylation, genomic instability, cancer, troubleshooting
MeSH terms: Base Sequence, Cloning, Molecular, DNA, Laboratories, Sulfites, Polymerase Chain Reaction, Sequence Analysis, DNA, DNA Methylation, Genomics

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DNA Methylation Detection: Bisulfite Genomic Sequencing Analysis

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