Open AccessImprovement of the Mutation-Discrimination Threshold for Rare Point Mutations by a Separation-Free Ligase Detection Reaction Assay Based on Fluorescence Resonance Energy Transfer
Author(s) -
Kenta Hagihara,
Kazuhiko Tsukagoshi,
Chinami Nakajima,
Shinsuke Esaki,
Masahiko Hashimoto
Publication year - 2016
Publication title -
analytical sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.392
H-Index - 73
eISSN - 1348-2246
pISSN - 0910-6340
DOI - 10.2116/analsci.32.367
Subject(s) - chemistry , förster resonance energy transfer , cyanine , fluorescence , alexa fluor , acceptor , point mutation , mutation , dna ligase , resonance (particle physics) , analytical chemistry (journal) , photochemistry , microbiology and biotechnology , dna , chromatography , biochemistry , atomic physics , optics , gene , physics , biology , condensed matter physics
We previously developed a separation-free ligase detection reaction assay based on fluorescence resonance energy transfer from a donor quantum dot to an acceptor fluorescent dye. This assay could successfully detect one cancer mutation among 10 wild-type templates. In the current study, the mutation-discrimination threshold was improved by one order of magnitude by replacing the original acceptor dye (Alexa Fluor 647) with another fluorescent dye (Cyanine 5) that was spectrally similar but more fluorescent.