Curr Protoc Mol Biol. 2010 Jul;Chapter 7:Unit 7.9.1-17.

Bisulfite sequencing of DNA.

Darst RP, Pardo CE, Ai L, Brown KD, Kladde MP.

Source

University of Florida College of Medicine, Gainesville, Florida, USA.

Abstract

Exact positions of 5-methylcytosine (m(5)C) on a single strand of DNA can be determined by bisulfite genomic sequencing (BGS). Treatment with bisulfite ion preferentially deaminates unmethylated cytosines, which are then converted to uracil upon desulfonation. Amplifying regions of interest from deaminated DNA and sequencing products cloned from amplicons permits determination of methylation at single-nucleotide resolution along single DNA molecules, which is not possible with other methylation analysis techniques. This unit describes a BGS technique suitable for most DNA sources, including formaldehyde-fixed tissue. Considerations for experimental design and common sources of error are discussed.

PMID:: 20583099; [PubMed - indexed for MEDLINE]
PMCID:: PMC3214597

Free PMC Article

Images from this publication.See all images (2) Free text

Publication Types, MeSH Terms, Substances, Grant Support

Publication Types

MeSH Terms

Substances

Grant Support

LinkOut - more resources

Full Text Sources

Other Literature Sources

COS Scholar Universe

Supplemental Content

Save items

Related citations in PubMed

Review The bisulfite genomic sequencing used in the analysis of epigenetic states, a technique in the emerging environmental genotoxicology research. [Mutat Res. 2008]
Review The bisulfite genomic sequencing used in the analysis of epigenetic states, a technique in the emerging environmental genotoxicology research.
Hayatsu H. Mutat Res. 2008 Jul-Aug; 659(1-2):77-82. Epub 2008 May 14.
High-speed conversion of cytosine to uracil in bisulfite genomic sequencing analysis of DNA methylation. [DNA Res. 2004]
High-speed conversion of cytosine to uracil in bisulfite genomic sequencing analysis of DNA methylation.
Shiraishi M, Hayatsu H. DNA Res. 2004 Dec 31; 11(6):409-15.
Bisulfite genomic sequencing: systematic investigation of critical experimental parameters. [Nucleic Acids Res. 2001]
Bisulfite genomic sequencing: systematic investigation of critical experimental parameters.
Grunau C, Clark SJ, Rosenthal A. Nucleic Acids Res. 2001 Jul 1; 29(13):E65-5.
Accelerated bisulfite-deamination of cytosine in the genomic sequencing procedure for DNA methylation analysis. [Nucleic Acids Symp Ser (Oxf). ...]
Accelerated bisulfite-deamination of cytosine in the genomic sequencing procedure for DNA methylation analysis.
Hayatsu H, Negishi K, Shiraishi M. Nucleic Acids Symp Ser (Oxf). 2004; (48):261-2.
Review Discovery of bisulfite-mediated cytosine conversion to uracil, the key reaction for DNA methylation analysis--a personal account. [Proc Jpn Acad Ser B Phys Biol ...]
Review Discovery of bisulfite-mediated cytosine conversion to uracil, the key reaction for DNA methylation analysis--a personal account.
Hayatsu H. Proc Jpn Acad Ser B Phys Biol Sci. 2008; 84(8):321-30.

See reviews... See all...

Related information

Related Citations
Calculated set of PubMed citations closely related to the selected article(s) retrieved using a word weight algorithm. Related articles are displayed in ranked order from most to least relevant, with the “linked from” citation displayed first.
Compound (MeSH Keyword)
PubChem chemical compound records that are classified under the same Medical Subject Headings (MeSH) controlled vocabulary as the current articles.
References for this PMC Article
Citation referenced in PubMed article. Only valid for PubMed citations that are also in PMC.
Substance (MeSH Keyword)
PubChem chemical substance (submitted) records that are classified under the same Medical Subject Headings (MeSH) controlled vocabulary as the current articles.
Free in PMC
Free full text articles in PMC

Recent activity

Clear Turn Off Turn On

Bisulfite sequencing of DNA.
Bisulfite sequencing of DNA.
Curr Protoc Mol Biol. 2010 Jul ;Chapter 7:Unit 7.9.1-17.

PubMed
Analysis of serine-threonine kinase specificity using arrayed positional scannin...
Analysis of serine-threonine kinase specificity using arrayed positional scanning peptide libraries.
Curr Protoc Mol Biol. 2010 Jul ;Chapter 18:Unit 18.14.

PubMed
Ultrastructural changes of cornea after ethanol ingestion in Otsuka Long-Evans T...
Ultrastructural changes of cornea after ethanol ingestion in Otsuka Long-Evans Tokushima fatty (OLETF) and Long-Evans Tokushima Otsuka (LETO) rats.
Graefes Arch Clin Exp Ophthalmol. 2010 Oct ;248(10):1457-66. Epub 2010 Jun 27 .

PubMed
Operational fidelity to an evidence-based HIV prevention intervention for people...
Operational fidelity to an evidence-based HIV prevention intervention for people living with HIV/AIDS.
J Prim Prev. 2010 Aug ;31(4):235-45.

PubMed
A guide to successful bioprospecting: informed by actinobacterial systematics.
A guide to successful bioprospecting: informed by actinobacterial systematics.
Antonie Van Leeuwenhoek. 2010 Aug ;98(2):119-42. Epub 2010 Jun 26 .

PubMed

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

PubMed

Result Filters

Display Settings:

Send to: