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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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2011
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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. |
format | Online Article Text |
id | pubmed-3130292 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT kimuramasayuki measurementoftelomerednacontentbydotblotanalysis AT avivabraham measurementoftelomerednacontentbydotblotanalysis |