Synthesis and evaluation of phosphorescent oligonucleotide probes for hybridisation assays

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dc.contributor.authorO'Sullivan, Paul J.
dc.contributor.authorBurke, Martina
dc.contributor.authorSoini, Aleski E.
dc.contributor.authorPapkovsky, Dmitri B.
dc.contributor.funderEnterprise Ireland
dc.date.accessioned2017-11-14T13:24:33Z
dc.date.available2017-11-14T13:24:33Z
dc.date.issued2002
dc.description.abstractMonofunctional, p-isothiocyanatophenyl-derivatives of platinum (II)-coproporphyrin-I (PtCP-NCS) were evaluated as phosphorescent labelling reagents for synthetic oligonucleotides containing a 3'- or 5'-amino modification. Synthesis and purification conditions were optimised to generate high yields and purity of PtCP-labelled oligonucleotide probes. Phosphorescent properties of the PtCP label have been shown to be largely unaffected by conjugation to oligonucleotides of various length, GC composition and label attachment site. 5'-PtCP-labelled oligonucleotides were shown to work efficiently as primers in a standard PCR. A dedicated 532 nm laser-based time-resolved fluorescence plate reader enabled highly sensitive detection of PtCP-labelled oligonucleotides and PCR products, both in solution and in agarose gels, with limits of detection in the order of 0.3 pM. A model system employing two complementary oligonucleotides labelled with PtCP and QSY(R) 7 dye (dark quencher) showed strong (similar to20-fold) and specific proximity quenching of PtCP label upon hybridisation in solution. The potential applications of PtCP-labelled probes in hybridisation assays were discussed.en
dc.description.sponsorshipEnterprise Ireland (IF/2001/008)
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.articleide114
dc.identifier.citationO’Sullivan, P. J., Burke, M., Soini, A. E. and Papkovsky, D. B. (2002) 'Synthesis and evaluation of phosphorescent oligonucleotide probes for hybridisation assays', Nucleic Acids Research, 30(21), e114 (7pp). doi: 10.1093/nar/gnf113en
dc.identifier.doi10.1093/nar/gnf113
dc.identifier.issn0305-1048
dc.identifier.issued21
dc.identifier.journaltitleNucleic Acids Researchen
dc.identifier.urihttps://hdl.handle.net/10468/5036
dc.identifier.volume30
dc.language.isoenen
dc.publisherOxford University Pressen
dc.relation.urihttps://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gnf113
dc.subjectTime-resolved fluorescenceen
dc.subjectNucleic acid hybridizationen
dc.subjectResonance energy transferen
dc.subjectEuropium chelateen
dc.subjectDNAen
dc.subjectCoproporphyrinen
dc.subjectPalladium(ii)en
dc.subjectFluorometryen
dc.subjectPrimersen
dc.subjectLabelsen
dc.titleSynthesis and evaluation of phosphorescent oligonucleotide probes for hybridisation assaysen
dc.typeArticle (peer-reviewed)en
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