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Measurement of telomere DNA content by dot blot analysis

Telomeres play a central role in human cancer, cardiovascular aging and possibly longevity. However, present methods to measure telomere length are fraught with shortcomings that limit their use. Here, we describe a novel method to measure the relative telomere DNA content by dot blot analysis. In e...

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Detalles Bibliográficos
Autores principales: Kimura, Masayuki, Aviv, Abraham
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3130292/
https://www.ncbi.nlm.nih.gov/pubmed/21525130
http://dx.doi.org/10.1093/nar/gkr235
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author Kimura, Masayuki
Aviv, Abraham
author_facet Kimura, Masayuki
Aviv, Abraham
author_sort Kimura, Masayuki
collection PubMed
description Telomeres play a central role in human cancer, cardiovascular aging and possibly longevity. However, present methods to measure telomere length are fraught with shortcomings that limit their use. Here, we describe a novel method to measure the relative telomere DNA content by dot blot analysis. In each dot, the DNA content is measured by a DNA stain (Dx) and the telomeric DNA content is measured with a telomeric probe (T). The T normalized for Dx (T/Dx) of each dot is a measure of telomere content. The method requires ∼20 ng of DNA per assay. Moreover, the T/Dx data are highly correlated linearly with mean telomere lengths derived from Southern blots of the terminal restriction fragments (r > 0.96, P < 0.0001). The method is also simple to use, has a relatively low interassay coefficient of variation (<6%), retains its precision in moderately degraded DNA and can be forged for high throughput analysis. The method might help researchers and clinicians alike in understanding risks for and extent of human diseases.
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spelling pubmed-31302922011-07-06 Measurement of telomere DNA content by dot blot analysis Kimura, Masayuki Aviv, Abraham Nucleic Acids Res Methods Online Telomeres play a central role in human cancer, cardiovascular aging and possibly longevity. However, present methods to measure telomere length are fraught with shortcomings that limit their use. Here, we describe a novel method to measure the relative telomere DNA content by dot blot analysis. In each dot, the DNA content is measured by a DNA stain (Dx) and the telomeric DNA content is measured with a telomeric probe (T). The T normalized for Dx (T/Dx) of each dot is a measure of telomere content. The method requires ∼20 ng of DNA per assay. Moreover, the T/Dx data are highly correlated linearly with mean telomere lengths derived from Southern blots of the terminal restriction fragments (r > 0.96, P < 0.0001). The method is also simple to use, has a relatively low interassay coefficient of variation (<6%), retains its precision in moderately degraded DNA and can be forged for high throughput analysis. The method might help researchers and clinicians alike in understanding risks for and extent of human diseases. Oxford University Press 2011-07 2011-04-27 /pmc/articles/PMC3130292/ /pubmed/21525130 http://dx.doi.org/10.1093/nar/gkr235 Text en © The Author(s) 2011. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Kimura, Masayuki
Aviv, Abraham
Measurement of telomere DNA content by dot blot analysis
title Measurement of telomere DNA content by dot blot analysis
title_full Measurement of telomere DNA content by dot blot analysis
title_fullStr Measurement of telomere DNA content by dot blot analysis
title_full_unstemmed Measurement of telomere DNA content by dot blot analysis
title_short Measurement of telomere DNA content by dot blot analysis
title_sort measurement of telomere dna content by dot blot analysis
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3130292/
https://www.ncbi.nlm.nih.gov/pubmed/21525130
http://dx.doi.org/10.1093/nar/gkr235
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