Rapid and Convenient Sample Preparation in a Single Tube Using Magnetic Beads for Fluorescence Detection of Single Nucleotide Variation Based on Oligonucleotide Ligation

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  • Masahiko Hashimoto
    Department of Chemical Engineering and Materials Science, Doshisha University
  • Chika Morimoto
    Department of Chemical Engineering and Materials Science, Doshisha University
  • Kenta Hagihara
    Department of Chemical Engineering and Materials Science, Doshisha University
  • Kazuhiko Tsukagoshi
    Department of Chemical Engineering and Materials Science, Doshisha University

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Abstract

<jats:title>Abstract</jats:title> <jats:p>We here describe a rapid and convenient sample preparation in a single tube for detecting single nucleotide variation based on oligonucleotide ligation assay (OLA) using a biotinylated primer and a set of four different fluorophore-labeled primers. Only one of the fluorophore-labeled primers that was fully complementary to a DNA template was ligated with the biotin-primer by DNA ligase. We isolated the ligated fragments that carried a biotin at the 3′-end by capture with streptavidin-coated magnetic beads. The distinct fluorescence signature of each ligation product coded the single nucleotide differences.</jats:p>

Journal

  • Chemistry Letters

    Chemistry Letters 41 (2), 135-137, 2012-01-28

    Oxford University Press (OUP)

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