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Sensitive fluorescence detection of nucleic acids based on isothermal circular strand-displacement polymerization reaction
Author(s) -
Qiuping Guo,
Xiaohai Yang,
Kemin Wang,
Weihong Tan,
Wei Li,
Hongxing Tang,
Huimin Li
Publication year - 2009
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/gkn1024
Subject(s) - polymerization , loop mediated isothermal amplification , primer (cosmetics) , dna , nucleic acid , multiple displacement amplification , biology , polymerase , chain reaction , nucleic acid thermodynamics , biophysics , recombinase polymerase amplification , fluorescence , polymerase chain reaction , microbiology and biotechnology , biochemistry , chemistry , photochemistry , polymer , base sequence , gene , physics , organic chemistry , quantum mechanics , dna extraction
Here we have developed a sensitive DNA amplified detection method based on isothermal strand-displacement polymerization reaction. This method takes advantage of both the hybridization property of DNA and the strand-displacement property of polymerase. Importantly, we demonstrate that our method produces a circular polymerization reaction activated by the target, which essentially allows it to self-detect. Functionally, this DNA system consists of a hairpin fluorescence probe, a short primer and polymerase. Upon recognition and hybridization with the target ssDNA, the stem of the hairpin probe is opened, after which the opened probe anneals with the primer and triggers the polymerization reaction. During this process of the polymerization reaction, a complementary DNA is synthesized and the hybridized target is displaced. Finally, the displaced target recognizes and hybridizes with another probe, triggering the next round of polymerization reaction, reaching a target detection limit of 6.4 x 10(-15) M.

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