Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Viterbo, David, Marchal, Astrid, Mosbach, Valentine, Poggi, Lucie, Vaysse-Zinkhöfer, Wilhelm, Richard, Guy-Franck
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2018
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
_version_ 1783493144609816576
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
work_keys_str_mv AT viterbodavid afastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT marchalastrid afastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT mosbachvalentine afastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT poggilucie afastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT vayssezinkhoferwilhelm afastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT richardguyfranck afastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT viterbodavid fastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT marchalastrid fastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT mosbachvalentine fastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT poggilucie fastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT vayssezinkhoferwilhelm fastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes
AT richardguyfranck fastsensitiveandcosteffectivemethodfornucleicaciddetectionusingnonradioactiveprobes