Show simple item record

dc.contributor.authorLi, Jianwei Jeffrey
dc.contributor.authorChu, Yizhuo
dc.contributor.authorLee, Benjamin Yi-Hung
dc.contributor.authorXie, Xiaoliang Sunney
dc.date.accessioned2010-02-12T18:25:00Z
dc.date.issued2008
dc.identifier.citationLi, Jianewi Jeffrey, Yizhuo Chu, Benjamin Yi-Hung Lee, and Xialong Sunney Xie. 2008. Enzymatic signal amplification of molecular beacons for sensitive DNA detection. Nucleic Acids Research 36(6): e36.en_US
dc.identifier.issn0305-1048en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:3637106
dc.description.abstractMolecular beacons represent a new family of fluorescent probes for nucleic acids, and have found broad applications in recent years due to their unique advantages over traditional probes. Detection of nucleic acids using molecular beacons has been based on hybridization between target molecules and molecular beacons in a 1:1 stoichiometric ratio. The stoichiometric hybridization, however, puts an intrinsic limitation on detection sensitivity, because one target molecule converts only one beacon molecule to its fluorescent form. To increase the detection sensitivity, a conventional strategy has been target amplification through polymerase chain reaction. Instead of target amplification, here we introduce a scheme of signal amplification, nicking enzyme signal amplification, to increase the detection sensitivity of molecular beacons. The mechanism of the signal amplification lies in target-dependent cleavage of molecular beacons by a DNA nicking enzyme, through which one target DNA can open many beacon molecules, giving rise to amplification of fluorescent signal. Our results indicate that one target DNA leads to cleavage of hundreds of beacon molecules, increasing detection sensitivity by nearly three orders of magnitude. We designed two versions of signal amplification. The basic version, though simple, requires that nicking enzyme recognition sequence be present in the target DNA. The extended version allows detection of target of any sequence by incorporating rolling circle amplification. Moreover, the extended version provides one additional level of signal amplification, bringing the detection limit down to tens of femtomolar, nearly five orders of magnitude lower than that of conventional hybridization assay.en_US
dc.description.sponsorshipChemistry and Chemical Biologyen_US
dc.language.isoen_USen_US
dc.publisherOxford University Pressen_US
dc.relation.isversionofhttp://dx.doi.org/10.1093/nar/gkn033en_US
dc.relation.hasversionhttp://bernstein.harvard.edu/papers/Jeff_Li_08.pdfen_US
dash.licenseMETA_ONLY
dc.titleEnzymatic Signal Amplification of Molecular Beacons for Sensitive DNA Detectionen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalNucleic Acids Researchen_US
dash.depositing.authorXie, Xiaoliang Sunney
dash.embargo.until10000-01-01
dc.identifier.doi10.1093/nar/gkn033*
dash.contributor.affiliatedChu, Yizhuo
dash.contributor.affiliatedXie, Xiaoliang


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record