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High Performance Liquid Chromatography Separation of Epigenetic Cytosine Variants

Epigenetic modifications enable cells to genetically respond to chemical inputs from environmental sources. These marks play a pivotal role in normal biological processes (e.g., differentiation, host defense and metabolic programs) but also contribute to the development of a wide variety of patholog...

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Autores principales: Roberts, Caroline E., Raner, Gregory M., Isaacs, Gary D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6526450/
https://www.ncbi.nlm.nih.gov/pubmed/31164555
http://dx.doi.org/10.3390/mps1020010
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author Roberts, Caroline E.
Raner, Gregory M.
Isaacs, Gary D.
author_facet Roberts, Caroline E.
Raner, Gregory M.
Isaacs, Gary D.
author_sort Roberts, Caroline E.
collection PubMed
description Epigenetic modifications enable cells to genetically respond to chemical inputs from environmental sources. These marks play a pivotal role in normal biological processes (e.g., differentiation, host defense and metabolic programs) but also contribute to the development of a wide variety of pathological conditions (e.g., cancer and Alzheimer’s disease). In particular, DNA methylation represents very stable epigenetic modification of cytosine bases that is strongly associated with a reduction in gene activity. Although High Performance Liquid Chromatography (HPLC) methodologies have been used to resolve methylated cytosine from unmodified cytosine bases, these represent only two of the five major cytosine analogs in the cell. Moreover, failure to resolve these other cytosine analogs might affect an accurate description of the cytosine methylation status in cells. In this present study, we determined the HPLC conditions required to separate the five cytosine analogs of the methylation/demethylation pathway. This methodology not only provides a means to analyze cytosine methylation as a whole, but it could also be used to more accurately calculate the methylation ratio from biological samples.
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spelling pubmed-65264502019-05-31 High Performance Liquid Chromatography Separation of Epigenetic Cytosine Variants Roberts, Caroline E. Raner, Gregory M. Isaacs, Gary D. Methods Protoc Technical Note Epigenetic modifications enable cells to genetically respond to chemical inputs from environmental sources. These marks play a pivotal role in normal biological processes (e.g., differentiation, host defense and metabolic programs) but also contribute to the development of a wide variety of pathological conditions (e.g., cancer and Alzheimer’s disease). In particular, DNA methylation represents very stable epigenetic modification of cytosine bases that is strongly associated with a reduction in gene activity. Although High Performance Liquid Chromatography (HPLC) methodologies have been used to resolve methylated cytosine from unmodified cytosine bases, these represent only two of the five major cytosine analogs in the cell. Moreover, failure to resolve these other cytosine analogs might affect an accurate description of the cytosine methylation status in cells. In this present study, we determined the HPLC conditions required to separate the five cytosine analogs of the methylation/demethylation pathway. This methodology not only provides a means to analyze cytosine methylation as a whole, but it could also be used to more accurately calculate the methylation ratio from biological samples. MDPI 2018-03-21 /pmc/articles/PMC6526450/ /pubmed/31164555 http://dx.doi.org/10.3390/mps1020010 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Technical Note
Roberts, Caroline E.
Raner, Gregory M.
Isaacs, Gary D.
High Performance Liquid Chromatography Separation of Epigenetic Cytosine Variants
title High Performance Liquid Chromatography Separation of Epigenetic Cytosine Variants
title_full High Performance Liquid Chromatography Separation of Epigenetic Cytosine Variants
title_fullStr High Performance Liquid Chromatography Separation of Epigenetic Cytosine Variants
title_full_unstemmed High Performance Liquid Chromatography Separation of Epigenetic Cytosine Variants
title_short High Performance Liquid Chromatography Separation of Epigenetic Cytosine Variants
title_sort high performance liquid chromatography separation of epigenetic cytosine variants
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6526450/
https://www.ncbi.nlm.nih.gov/pubmed/31164555
http://dx.doi.org/10.3390/mps1020010
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