Synthesis and evaluation of phosphorescent oligonucleotide probes for hybridisation assays (2002).

Content Provider	CiteSeerX
Author	O.'Sullivan, Paul J. Burke, Martina Soini, Aleksi E. Papkovsky, Dmitri B.
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 modi®cation. Synthesis and puri®cation con-ditions 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 compos-ition and label attachment site. 5¢-PtCP-labelled oligonucleotides were shown to work ef®ciently as primers in a standard PCR. A dedicated 532 nm laser-based time-resolved ¯uorescence plate reader enabled highly sensitive detection of PtCP-labelled oligonucleotides and PCR products, both in solu-tion 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 QSYâ 7 dye (dark quencher) showed strong (~20-fold) and speci®c proximity quenching of PtCP label upon hybridisation in solution. The potential applications of PtCP-labelled probes in hybridisa-tion assays were discussed.
File Format	PDF
Publisher Date	2002-01-01
Access Restriction	Open
Subject Keyword	Phosphorescent Oligonucleotide Probe Hybridisation Assay Ptcp Label Ptcp-labelled Oligonucleotides Pcr Product Amino Modi Cation Agarose Gel High Yield Puri Cation Con-ditions Sensitive Detection Phosphorescent Property Synthetic Oligonucleotides Complementary Oligonucleotides Gc Compos-ition Ptcp-labelled Probe Hybridisa-tion Assay Various Length Model System Label Attachment Site Speci Proximity Quenching Standard Pcr Dark Quencher Phosphorescent Labelling Reagent Potential Application
Content Type	Text