Label-Free in Situ Monitoring of the DNA Hybridization Chain Reaction by Using Sequence-Selective Minor-Groove-Binding Fluorophores

被引:3
|
作者
Sakamoto, Takashi [1 ]
Yamada, Rikuto [1 ]
机构
[1] Wakayama Univ, Fac Syst Engn, 930 Sakaedani, Wakayama 6408510, Japan
关键词
DNA structures; fluorescent probes; hybridization chain reaction; in situ detection; minor-groove-binding fluorophores; MESSENGER-RNA; PROBE; AMPLIFICATION; DESIGN;
D O I
10.1002/cbic.201800738
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A new label-free in situ monitoring system for the hybridization chain reaction (HCR) based on DNA minor-groove-binding fluorophores [Hoechst 33258 (Hoe) or quinone cyanine-dithiazole (QCy-DT)] has been developed. Use of two unmodified hairpin oligodeoxyribonucleotides containing incomplete double-stranded AATT sequences enabled target-dependent formation of probe binding sitesthat is, AATT double strandin the HCR product, together with fluorescence enhancement of minor-groove-binding fluorophores in situ. This system allows target DNA to be detected through the fluorescence enhancement of Hoe and QCy-DT in real time and in situ. Further development of a label-free, isothermal detection system might provide a cost-effective and user-friendly method for nucleic acid detection.
引用
收藏
页码:1242 / 1245
页数:4
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