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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...

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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
Descripción
Sumario: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.