Response surface methodology to optimize peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) in Saccharomyces cerevisiae

• Type: Article in International Scientific Journal
• Date: 2017

Authors

Rita S. Santos (Author) FEUP; Carolina C. Lima (Author) Other; Daniel Carvalho (Author) Other; Francisco Meireles (Author) Other; Nuno Guimarães (Author) FEUP; Nuno F. Azevedo (Author) FEUP

Journal

Title: LWT - Food Science and Technology

Vol. 80

Pages: 27-31

ISSN: 0023-6438

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 3 Citations

Scopus - 3 Citations

Associated Projects

Coded-FISH - Desenvolvimento de um novo método de RING FISH codificado por cores: uma ferramenta inovadora para a deteção de patogénios resistentes a antibióticos

LEPABE - Laboratório de Engenharia de Processos, Ambiente, Biotecnologia e Energia

Other information

Authenticus ID: P-00M-GGC

DOI: 10.1016/j.lwt.2017.01.077

Abstract (EN): Saccharomyces cerevisiae (S. cerevisiae) is a crucial fermenting microorganism for the beer, wine and bread industry. As such, an accurate and rapid method for its identification and monitoring is required. In here, a peptide nucleic acid (PNA) probe was first designed to target S. cerevisiae by fluorescence in situ hybridization (FISH). This PNA-FISH method was then systematically optimized, employing response surface methodology (RSM). The interaction between the critical hybridization temperature, time and formamide concentration and their effect on the FISH efficiency were modelled. The model predicted optimal 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 a valuable tool to optimize and better understand the dynamics of yeast FISH, which can impact the performance evaluation of related fermentation processes.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 5