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Split Spinach Aptamer for Highly Selective Recognition of DNA and RNA at Ambient Temperatures
Author(s) -
Kikuchi Nanami,
Kolpashchikov Dmitry M.
Publication year - 2016
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.201600323
Subject(s) - nucleic acid , aptamer , analyte , fluorescence , rna , dna , chemistry , spinach , combinatorial chemistry , molecular beacon , nucleic acid structure , biochemistry , biophysics , oligonucleotide , chromatography , biology , microbiology and biotechnology , gene , physics , quantum mechanics
Split spinach aptamer (SSA) probes for fluorescent analysis of nucleic acids were designed and tested. In SSA design, two RNA or RNA/DNA strands hybridized to a specific nucleic acid analyte and formed a binding site for low‐fluorescent 3,5‐difluoro‐4‐hydroxybenzylidene imidazolinone (DFHBI) dye, which resulted in up to a 270‐fold increase in fluorescence. The major advantage of the SSA over state‐of‐the art fluorescent probes is high selectivity: it produces only background fluorescence in the presence of a single‐base‐mismatched analyte, even at room temperature. SSA is therefore a promising tool for label‐free analysis of nucleic acids at ambient temperatures.