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Author(s): Rita S. Santos
Carolina C. Lima
Daniel Carvalho
Francisco Meireles
Nuno Guimarães
Nuno F. Azevedo
Title: Response surface methodology to optimize peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) in Saccharomyces cerevisiae
Issue Date: 2017
Abstract: Saccharomyces cerevisiae (S. cerevisiae) is a crucial fermenting microorganism for the beer, wine andbread industry. As such, an accurate and rapid method for its identification and monitoring is required. Inhere, a peptide nucleic acid (PNA) probe was first designed to target S. cerevisiae by fluorescence in situhybridization (FISH). This PNA-FISH method was then systematically optimized, employing responsesurface methodology (RSM). The interaction between the critical hybridization temperature, time andformamide concentration and their effect on the FISH efficiency were modelled. The model predictedoptimal fluorescence intensity upon hybridization at 53.9 C, during 57.8 min and using 43.8% (v/v)formamide in the hybridization buffer, which was experimentally confirmed. RSM showed to be avaluable tool to optimize and better understand the dynamics of yeast FISH, which can impact theperformance evaluation of related fermentation processes.
Related Information: info:eu-repo/grantAgreement/FCT - Fundação para a Ciência e Tecnologia/P2020|COMPETE - Projetos em Todos os Domínios Científicos/PTDC/DTP-PIC/4562/2014 - POCI-01-0145-FEDER-016678/Desenvolvimento de um novo método de RING FISH codificado por cores: uma ferramenta inovadora para a deteção de patogénios resistentes a antibióticos/Coded-FISH
info:eu-repo/grantAgreement/FCT - Fundação para a Ciência e Tecnologia/Projetos Estratégicos/UID/EQU/00511/2013 - POCI-01-0145-FEDER-006939/Laboratório de Engenharia de Processos, Ambiente, Biotecnologia e Energia/LEPABE
Document Type: Artigo em Revista Científica Internacional
Rights: restrictedAccess
Appears in Collections:FEUP - Artigo em Revista Científica Internacional

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