Evaluation of the phosphorescent palladium(II)-coproporphyrin labels in separation-free hybridization assays.
Palladium(II)-coproporphyrin label and a set of corresponding monofunctional labeling reagents with different linker arms were evaluated for labeling of oligonucleotides and subsequent use in hybridization assays. The properties of resulting oligonucleotide probes including phosphorescence spectra, quantum yields, lifetimes, and labeling yields were examined as functions of the label and oligonucleotide structures. Upon hybridization with complementary sequences bearing dabcyl, QSY-7, and rhodamine green dyes, the probes displayed strong quenching due to close proximity effects. Intensity and lifetime changes of the phosphorescence, distance, and temperature dependences were investigated in detail. The potential of the new label and probes for sensitive and separation-free hybridization assays was discussed.
Burke
Martina
M
Biochemistry Department/ABCRF, University College Cork, Lee Maltings, Cork, Ireland.
O'Sullivan
Paul J
PJ
Soini
Aleksi E
AE
Berney
Helen
H
Papkovsky
Dmitri B
DB
D001681:Biological Assay; D003306:Coproporphyrins / Q000737:chemistry; D008163:Luminescent Measurements; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides; D010165:Palladium / Q000737:chemistry; D013053:Spectrophotometry; D013194:Staining and Labeling
D003306:Coproporphyrins; D009841:Oligonucleotides; D010165:Palladium