Synthesis and evaluation of phosphorescent oligonucleotide probes for hybridisation assays

Show simple item record O'Sullivan, Paul J. Burke, Martina Soini, Aleski E. Papkovsky, Dmitri B. 2017-11-14T13:24:33Z 2017-11-14T13:24:33Z 2002
dc.identifier.citation O’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/gnf113 en
dc.identifier.volume 30
dc.identifier.issued 21
dc.identifier.issn 0305-1048
dc.identifier.doi 10.1093/nar/gnf113
dc.description.abstract Monofunctional, 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.sponsorship Enterprise Ireland (IF/2001/008)
dc.format.mimetype application/pdf en
dc.language.iso en en
dc.publisher Oxford University Press en
dc.subject Time-resolved fluorescence en
dc.subject Nucleic acid hybridization en
dc.subject Resonance energy transfer en
dc.subject Europium chelate en
dc.subject DNA en
dc.subject Coproporphyrin en
dc.subject Palladium(ii) en
dc.subject Fluorometry en
dc.subject Primers en
dc.subject Labels en
dc.title Synthesis and evaluation of phosphorescent oligonucleotide probes for hybridisation assays en
dc.type Article (peer-reviewed) en
dc.internal.authorcontactother Dmitri Papkovsky, Biochemistry & Cell Biology, University College Cork, Cork, Ireland. +353-21-490-3000 Email: en
dc.internal.availability Full text available en
dc.description.version Published Version en
dc.contributor.funder Enterprise Ireland
dc.description.status Peer reviewed en
dc.identifier.journaltitle Nucleic Acids Research en
dc.internal.IRISemailaddress en
dc.identifier.articleid e114

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