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A fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes
Nucleic acid detection and quantification using a labeled DNA probe is a very common molecular biology procedure. Here, we describe a new method, based on commonly used laboratory solutions, for nucleic acid hybridization and detection with digoxigenin-labeled DNA probes. The protocol described is f...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6994052/ https://www.ncbi.nlm.nih.gov/pubmed/32161800 http://dx.doi.org/10.1093/biomethods/bpy006 |
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author | Viterbo, David Marchal, Astrid Mosbach, Valentine Poggi, Lucie Vaysse-Zinkhöfer, Wilhelm Richard, Guy-Franck |
author_facet | Viterbo, David Marchal, Astrid Mosbach, Valentine Poggi, Lucie Vaysse-Zinkhöfer, Wilhelm Richard, Guy-Franck |
author_sort | Viterbo, David |
collection | PubMed |
description | Nucleic acid detection and quantification using a labeled DNA probe is a very common molecular biology procedure. Here, we describe a new method, based on commonly used laboratory solutions, for nucleic acid hybridization and detection with digoxigenin-labeled DNA probes. The protocol described is faster, more sensitive and much cheaper than a standard protocol using commercial solutions. Comparison with a classical radioactive detection method shows that the latter exhibits less background and shows a greater linear response. Hence, the proposed protocol may be routinely performed for qualitative detection of nucleic acid, but when precise signal quantitation needs to be obtained, radioactive probe hybridization associated to phosphorimaging technology is more reliable. |
format | Online Article Text |
id | pubmed-6994052 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-69940522020-03-11 A fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes Viterbo, David Marchal, Astrid Mosbach, Valentine Poggi, Lucie Vaysse-Zinkhöfer, Wilhelm Richard, Guy-Franck Biol Methods Protoc Methods Manuscript Nucleic acid detection and quantification using a labeled DNA probe is a very common molecular biology procedure. Here, we describe a new method, based on commonly used laboratory solutions, for nucleic acid hybridization and detection with digoxigenin-labeled DNA probes. The protocol described is faster, more sensitive and much cheaper than a standard protocol using commercial solutions. Comparison with a classical radioactive detection method shows that the latter exhibits less background and shows a greater linear response. Hence, the proposed protocol may be routinely performed for qualitative detection of nucleic acid, but when precise signal quantitation needs to be obtained, radioactive probe hybridization associated to phosphorimaging technology is more reliable. Oxford University Press 2018-06-25 /pmc/articles/PMC6994052/ /pubmed/32161800 http://dx.doi.org/10.1093/biomethods/bpy006 Text en © The Author(s) 2018. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Methods Manuscript Viterbo, David Marchal, Astrid Mosbach, Valentine Poggi, Lucie Vaysse-Zinkhöfer, Wilhelm Richard, Guy-Franck A fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes |
title | A fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes |
title_full | A fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes |
title_fullStr | A fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes |
title_full_unstemmed | A fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes |
title_short | A fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes |
title_sort | fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes |
topic | Methods Manuscript |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6994052/ https://www.ncbi.nlm.nih.gov/pubmed/32161800 http://dx.doi.org/10.1093/biomethods/bpy006 |
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