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DNA Methylation Detection: Bisulfite Genomic Sequencing Analysis
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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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“Bisulfite sequencing is used to identify methylated DNA (5mC) by converting all unmethylated cytosine bases to uracil through the addition of a bisulfate agent such as sodium bisulfate.”
From The Epigenetics of Anxiety Pathophysiology: A DNA Methylation and Histone Modification Focused Review (2023) · cites this paper as Li and Tollefsbol, 2011
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Metadata
- Authors
- Li, Y., & Tollefsbol, T. O.
- Affiliations
- University of Alabama at Birmingham (US)
- Corresponding author
- 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
- 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
- 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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