Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution

Bisulfite sequencing measures absolute levels of DNA methylation at single-nucleotide resolution, providing a robust platform for molecular diagnostics. Here, we optimize bisulfite sequencing for genome-scale analysis of clinical samples. Specifically, we outline how restriction digestion targets bi...

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Autores principales: Gu, Hongcang, Bock, Christoph, Mikkelsen, Tarjei S., Jäger, Natalie, Smith, Zachary D., Tomazou, Eleni, Gnirke, Andreas, Lander, Eric S., Meissner, Alexander
Formato: Texto
Lenguaje:English
Publicado: 2010
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2860480/
https://www.ncbi.nlm.nih.gov/pubmed/20062050
http://dx.doi.org/10.1038/nmeth.1414
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author Gu, Hongcang
Bock, Christoph
Mikkelsen, Tarjei S.
Jäger, Natalie
Smith, Zachary D.
Tomazou, Eleni
Gnirke, Andreas
Lander, Eric S.
Meissner, Alexander
author_facet Gu, Hongcang
Bock, Christoph
Mikkelsen, Tarjei S.
Jäger, Natalie
Smith, Zachary D.
Tomazou, Eleni
Gnirke, Andreas
Lander, Eric S.
Meissner, Alexander
author_sort Gu, Hongcang
collection PubMed
description Bisulfite sequencing measures absolute levels of DNA methylation at single-nucleotide resolution, providing a robust platform for molecular diagnostics. Here, we optimize bisulfite sequencing for genome-scale analysis of clinical samples. Specifically, we outline how restriction digestion targets bisulfite sequencing to hotspots of epigenetic regulation; we show that 30ng of DNA are sufficient for genome-scale analysis; we demonstrate that our protocol works well on formalin-fixed, paraffin-embedded (FFPE) samples; and we describe a statistical method for assessing significance of altered DNA methylation patterns.
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spelling pubmed-28604802010-08-01 Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution Gu, Hongcang Bock, Christoph Mikkelsen, Tarjei S. Jäger, Natalie Smith, Zachary D. Tomazou, Eleni Gnirke, Andreas Lander, Eric S. Meissner, Alexander Nat Methods Article Bisulfite sequencing measures absolute levels of DNA methylation at single-nucleotide resolution, providing a robust platform for molecular diagnostics. Here, we optimize bisulfite sequencing for genome-scale analysis of clinical samples. Specifically, we outline how restriction digestion targets bisulfite sequencing to hotspots of epigenetic regulation; we show that 30ng of DNA are sufficient for genome-scale analysis; we demonstrate that our protocol works well on formalin-fixed, paraffin-embedded (FFPE) samples; and we describe a statistical method for assessing significance of altered DNA methylation patterns. 2010-01-10 2010-02 /pmc/articles/PMC2860480/ /pubmed/20062050 http://dx.doi.org/10.1038/nmeth.1414 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Gu, Hongcang
Bock, Christoph
Mikkelsen, Tarjei S.
Jäger, Natalie
Smith, Zachary D.
Tomazou, Eleni
Gnirke, Andreas
Lander, Eric S.
Meissner, Alexander
Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution
title Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution
title_full Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution
title_fullStr Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution
title_full_unstemmed Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution
title_short Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution
title_sort genome-scale dna methylation mapping of clinical samples at single-nucleotide resolution
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2860480/
https://www.ncbi.nlm.nih.gov/pubmed/20062050
http://dx.doi.org/10.1038/nmeth.1414
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