RNA-FISH Technique: Probe Cocktails vs Single Probes

• Autor(a) principal: Vieira, Ricardo
• Data de Publicação: 2016
• Outros Autores: Pereira, António, Candeias, António, Caldeira, A. Teresa, González-Pérez, Marina
• Tipo de documento: Artigo de conferência
• Idioma: por
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10174/19481
• topic: Fluorescence In Situ Hybridization; Probe cocktails; RNA-FISH; Rhodotorula sp

RNA-Fluorescence In Situ Hybridization (RNA-FISH) enables to analyze and visualize the microorganisms of interest within microbial communities in their natural environments by fluorescent labelling of specific RNA sequences. Poor accessibility or low content of the RNA target region can cause false positives/negatives due to low fluorescence of the cells. To reduce the chances of this occurring, probe cocktails – i.e. mixture of several probes that hybridize to different regions of the target RNA- has been proposed as an alternative to single probes use for increasing the Fluorescence Intensities (FI). However, is this really a good solution? The key finding of this work was that the use of probe cocktails is not always a good solution for maximizing the FI as at high concentrations the single probe EUK516-6 FAM yielded higher FI than the probe cocktails.