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The quenching of electrochemiluminescence upon oligonucleotide hybridization
Author(s) -
Spehar AnnaMaria,
Koster Sander,
Kulmala Sakari,
Verpoorte Elisabeth,
de Rooij Nico,
KoudelkaHep Milena
Publication year - 2004
Publication title -
luminescence
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.428
H-Index - 45
eISSN - 1522-7243
pISSN - 1522-7235
DOI - 10.1002/bio.786
Subject(s) - electrochemiluminescence , quenching (fluorescence) , electron transfer , förster resonance energy transfer , luminescence , chemistry , oligonucleotide , excited state , fluorescence , photochemistry , resonance (particle physics) , excitation , analytical chemistry (journal) , electrode , materials science , atomic physics , physics , dna , biochemistry , optoelectronics , chromatography , quantum mechanics
Many genomic assays rely on a distance‐dependent interaction between luminescent labels, such as luminescence quenching or resonance energy transfer. We studied the interaction between electrochemically excited Ru(bpy) 3 2+ and Cy5 in a hybridization assay on a chip. The 3′ end of an oligonucleotide was labelled with Ru(bpy) 3 2+ and the 5′ end of a complementary strand with Cy5. Upon the hybridization, the electrochemiluminescence (ECL) of Ru(bpy) 3 2+ was efciently quenched by Cy5 with a sensitivity down to 30 nmol/L of the Cy5‐labelled complementary strand. The quenching efciency is calculated to be 78%. A similar phenomenon was observed in a comparative study using laser‐excitation of Ru(bpy) 3 2+ . The hybridization with the non‐labelled complementary or labelled non‐complementary strand did not change the intensity of the ECL signal. Resonance energy transfer, electron transfer and static quenching mechanisms are discussed. Our results suggest that static quenching and/or electron transfer are the most likely quenching mechanisms. Copyright © 2004 John Wiley & Sons, Ltd.