Improvement of the Mutation-Discrimination Threshold for Rare Point Mutations by a Separation-Free Ligase Detection Reaction Assay Based on Fluorescence Resonance Energy Transfer

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  • HAGIHARA Kenta
    Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University
  • TSUKAGOSHI Kazuhiko
    Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University
  • NAKAJIMA Chinami
    Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University
  • ESAKI Shinsuke
    Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University
  • HASHIMOTO Masahiko
    Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University

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Abstract

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.

Journal

  • Analytical Sciences

    Analytical Sciences 32 (3), 367-370, 2016

    The Japan Society for Analytical Chemistry

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