title: Fluorescent-labeled oligonucleotide probes: detection of hybrid formation in solution by fluorescence polarization spectroscopy.
abstract: Fluorescein-labeled oligonucleotides as DNA-probes were synthesized and used to monitor hybrid formation, namely to detect DNA or oligonucleotide sequence in solution. The introduction of fluorescein to oligonucleotides was carried out by oxidation of a hydrogen phosphonate linkage with ethylenediamine or hexamethylenediamine as a tether and by a subsequent labeling of the primary amine moiety by FITC. Fluorescence anisotropy, r, was adopted as an index to monitor the behavior of F-probe in solution. An increase in the anisotropy was observed upon an increase in the chain-length of F-probe. When F-Probe formed a hybrid with its complementary oligonucleotide in solution, the r value increased compared to that of F-Probe itself. These observations clearly indicate that measurements of r in solution will readily lead to the monitoring of the presence of a hybrid in solution. Consequently, it is promising to detect a certain nucleic acid sequence in solution using fluorescent-labeled oligonucleotides.
authors.affiliation: Department of Polymer Science and Engineering, Kyoto Institute of Technology, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Murakami
authors.forename: M
authors.initials: M
authors.lastname: Nakaura
authors.forename: Y
authors.initials: Y
authors.lastname: Nakatsuji
authors.forename: S
authors.initials: S
authors.lastname: Nagahara
authors.forename: Q
authors.initials: Q
authors.lastname: Tran-Cong
authors.forename: K
authors.initials: K
authors.lastname: Makino
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004277:DNA, Single-Stranded / Q000378:metabolism*; D019793:Fluorescein; D005452:Fluoresceins / Q000737:chemistry*; D005454:Fluorescence Polarization; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000138:chemical synthesis / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D013696:Temperature
chemical_list: D004277:DNA, Single-Stranded; D005452:Fluoresceins; D015345:Oligonucleotide Probes; D019793:Fluorescein