A fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes

David Viterbo (1) , Astrid Marchal (1) , Valentine Mosbach (1) , Lucie Poggi (1, 2, 3) , Wilhelm Vaysse-Zinkhöfer (1, 3) , Guy-Franck Richard (1)

1 Dynamique du Génome - Dynamics of the genome
2 Collège Doctoral
3 SANOFI Recherche

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.

Mots clés: nucleic acid hybridization; nucleic acid quantification; digoxigenin-labeled probe

Domaines: Sciences du Vivant [q-bio]

David Viterbo, Astrid Marchal, Valentine Mosbach, Lucie Poggi, Wilhelm Vaysse-Zinkhöfer, et al.. A fast, sensitive and cost-effective method for nucleic acid detection using non-radioactive probes. Biology Methods and Protocols, 2018, ⟨10.1093/biomethods/bpy006⟩. ⟨pasteur-02864565⟩