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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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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. |
format | Online Article Text |
id | pubmed-6526450 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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