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Reactive Quantum Dot-Based FRET Systems

  • Oligonucleotide-templated reactions (OTRs) between two reactive hybridization probes allow for the detection of a DNA or RNA of interest by exploiting the target molecule as a catalyst of chemical reactions. The product of such a reaction commonly exhibits distinct fluorescence properties and can be detected by the means of fluorescence spectroscopy. The vast majority of OTR systems utilize organic dyes as fluorescent reporters. However, the use of brighter emitters, such as semiconductor quantum dots (QDs), has potential to improve the sensitivity of detection by providing brighter signals and permitting the use of probes at very low concentrations. Here we report an RNA-templated reaction between two fluorescently labeled peptide nucleic acid (PNA)-based probes, which proceeds on the surface of a QD. The QD-Bound PNA probe bears a cysteine functionality, while the other PNA is functionalized with an organic dye as a thioester. OTR between these probes proceeds through a transfer ofOligonucleotide-templated reactions (OTRs) between two reactive hybridization probes allow for the detection of a DNA or RNA of interest by exploiting the target molecule as a catalyst of chemical reactions. The product of such a reaction commonly exhibits distinct fluorescence properties and can be detected by the means of fluorescence spectroscopy. The vast majority of OTR systems utilize organic dyes as fluorescent reporters. However, the use of brighter emitters, such as semiconductor quantum dots (QDs), has potential to improve the sensitivity of detection by providing brighter signals and permitting the use of probes at very low concentrations. Here we report an RNA-templated reaction between two fluorescently labeled peptide nucleic acid (PNA)-based probes, which proceeds on the surface of a QD. The QD-Bound PNA probe bears a cysteine functionality, while the other PNA is functionalized with an organic dye as a thioester. OTR between these probes proceeds through a transfer of the organic dye to the QD and can be conveniently monitored via fluorescence resonance energy transfer (FRET) from the QD to the Cy5. The reaction was performed in a conventional fluorescence microplate reader and permits the detection of RNA in the picomolar range.zeige mehrzeige weniger

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  • OZ_Protocol 2020_Reactive Quantum Dot-Based FRET Systems forTarget-Catalyzed Detection of RNA.pdf
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Metadaten
Autor*innen:O. Zavoiura, Ute Resch-GengerORCiD, O. Seitz
Dokumenttyp:Zeitschriftenartikel
Veröffentlichungsform:Verlagsliteratur
Sprache:Englisch
Titel des übergeordneten Werkes (Englisch):Methods in Molecular Biology
Jahr der Erstveröffentlichung:2020
Organisationseinheit der BAM:1 Analytische Chemie; Referenzmaterialien
1 Analytische Chemie; Referenzmaterialien / 1.2 Biophotonik
Verlag:Springer Sience+Business Media
Jahrgang/Band:2105
Erste Seite:187
Letzte Seite:198
Freie Schlagwörter:Assay; Bioconjugation; Click chemistry; Dye; FRET; Fluorescence; Fluorescence spectroscopy; Nano; Particle quantum dot; Peptide nucleic acid; Quantum dots; RNA; RNA-templated reactions; Sensor; Synthesis
Themenfelder/Aktivitätsfelder der BAM:Material
Umwelt
Umwelt / Sensorik
DOI:10.1007/978-1-0716-0243-0_11
Verfügbarkeit des Dokuments:Datei im Netzwerk der BAM verfügbar ("Closed Access")
Datum der Freischaltung:16.03.2020
Referierte Publikation:Ja
Datum der Eintragung als referierte Publikation:16.03.2020
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