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Author Torimura, Masaki ♦ Kurata, Shinya ♦ Yamada, Kazutaka ♦ Yokomaku, Toyokazu ♦ Kamagata, Yoichi ♦ Kanagawa, Takahiro ♦ Kurane, Ryuichiro
Source J-STAGE
Content type Text
Publisher The Japan Society for Analytical Chemistry
Language English
Abstract Fluorescently labeled oligonucleotide probes have been widely used in biotechnology, and fluorescence quenching by the interaction between the dyes and a nucleobase has been pointed out. This quenching causes big problem in analytical methods, but is useful in some other cases. Therefore, it is necessary to estimate the fluorescence quenching intensity under various conditions. We focused on the redox properties of some commercially available fluorescent dyes, and investigated dye-nucleotide interactions between a free dye and a nucleotide in aqueous solution by electrochemical and spectroscopic techniques. Our results suggested that the quenching was accompanied by photoinduced electron transfer between a thermodynamically quenchable excited dye and a specific base. Several kinds of fluorescent dyes labeled to the 5′-end of oligonucleotide $C_{10}T_{6}$ were prepared, and their quenching ratios compared upon hybridization with the complementary oligonucleotide $A_{6}G_{10}.$ The quenching was completely reversible and their efficiencies depended on the attached fluorophore types. The fluorescence of 5-FAM, BODIPY FL or TAMRA-modified probe was strongly quenched by hybridization.
ISSN 09106340
Learning Resource Type Article
Publisher Date 2001-01-01
e-ISSN 13482246
Journal Analytical Sciences(analsci)
Volume Number 17
Issue Number 1
Page Count 6
Starting Page 155
Ending Page 160


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