title: Improvement of the Mutation-Discrimination Threshold for Rare Point Mutations by a Separation-Free Ligase Detection Reaction Assay Based on Fluorescence Resonance Energy Transfer.
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.
authors.affiliation: Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University.
authors.forename: Kenta
authors.initials: K
authors.lastname: Hagihara
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Tsukagoshi
authors.forename: Chinami
authors.initials: C
authors.lastname: Nakajima
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Esaki
authors.forename: Masahiko
authors.initials: M
authors.lastname: Hashimoto
mesh_terms: D002232:Carbocyanines / Q000737:chemistry*; D015179:Colorectal Neoplasms / Q000235:genetics; D004247:DNA / Q000235:genetics; D011088:DNA Ligases / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods / Q000592:standards; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods* / Q000592:standards; D005456:Fluorescent Dyes / Q000737:chemistry*; D011905:Genes, ras / Q000235:genetics; D019073:HT29 Cells; D006801:Humans; D022521:Ligase Chain Reaction / Q000379:methods* / Q000592:standards; D008025:Ligases / Q000235:genetics*; D017354:Point Mutation; D045663:Quantum Dots; D012680:Sensitivity and Specificity
chemical_list: D002232:Carbocyanines; D005456:Fluorescent Dyes; C085321:cyanine dye 5; D004247:DNA; D008025:Ligases; D011088:DNA Ligases