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Label‐Free in Situ Monitoring of the DNA Hybridization Chain Reaction by Using Sequence‐Selective Minor‐Groove‐Binding Fluorophores
Author(s) -
Sakamoto Takashi,
Yamada Rikuto
Publication year - 2019
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800738
Subject(s) - minor groove , dna , in situ , chain reaction , chemistry , sequence (biology) , biophysics , stereochemistry , combinatorial chemistry , biochemistry , biology , photochemistry , organic chemistry
A new label‐free in situ monitoring system for the hybridization chain reaction (HCR) based on DNA minor‐groove‐binding fluorophores [Hoechst 33258 (Hoe) or quinone cyanine‐dithiazole (QCy‐DT)] has been developed. Use of two unmodified hairpin oligodeoxyribonucleotides containing incomplete double‐stranded AATT sequences enabled target‐dependent formation of probe binding sites—that is, AATT double strand—in the HCR product, together with fluorescence enhancement of minor‐groove‐binding fluorophores in situ. This system allows target DNA to be detected through the fluorescence enhancement of Hoe and QCy‐DT in real time and in situ. Further development of a label‐free, isothermal detection system might provide a cost‐effective and user‐friendly method for nucleic acid detection.