Please use this identifier to cite or link to this item: https://hdl.handle.net/10216/104675
Author(s): Sílvia Fontenete
Joana Barros
Pedro Madureira
Céu Figueiredo
Jesper Wengel
Nuno Filipe Azevedo
Title: Mismatch discrimination in fluorescent in situ hybridization using different types of nucleic acids
Issue Date: 2015
Abstract: In the past few years, several researchers have focused their attention on nucleic acid mimics due to the increasing necessity of developing a more robust recognition of DNAor RNA sequences. Fluorescence in situ hybridization (FISH)is an example of a method where the use of these novel nucleicacid monomers might be crucial to the success of the analysis.To achieve the expected accuracy in detection, FISH probesshould have high binding affinity towards their complementary strands and discriminate effectively the noncomplementary strands. In this study, we investigate the effect of differentchemical modifications in fluorescent probes on their abilityto successfully detect the complementary target and discriminate the mismatched base pairs by FISH. To our knowledge,this paper presents the first study where this analysis is performed with different types of FISH probes directly in biological targets, Helicobacter pylori and Helicobacteracinonychis. This is also the first study where unlockednucleic acids (UNA) were used as chemistry modification inoligonucleotides for FISH methodologies. The effectivenessin detecting the specific target and in mismatch discriminationappears to be improved using locked nucleic acids (LNA)/2′-O-methyl RNA (2′OMe) or peptide nucleic acid (PNA) incomparison to LNA/DNA, LNA/UNA, or DNA probes.Further, the use of LNA modifications together with 2′OMemonomers allowed the use of shorter fluorescent probes andincreased the range of hybridization temperatures at whichFISH would work.
URI: https://hdl.handle.net/10216/104675
Related Information: info:eu-repo/grantAgreement/FCT - Fundação para a Ciência e Tecnologia/Projectos de Investigação Clínica/PIC/IC/82815/2007/Desenvolvimento e aplicação de mímicos de DNA para a rápida identificação de microrganismos patogénicos/DNA mimics
info:eu-repo/grantAgreement/Autoriadade de Gestão do Programa Operacional Regional do Norte/Programas Integrados de IC&DT/NORTE-07-0124-FEDER-000025/(Bio) Chemical Engineering: Multi-Scale Approaches for Sustainable Environment and Health/LEPAE/CEFT - RL2
Document Type: Artigo em Revista Científica Internacional
Rights: restrictedAccess
Appears in Collections:FEUP - Artigo em Revista Científica Internacional

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