title: Europium-labeled oligonucleotide hybridization probes: preparation and properties.
abstract: A chemical method for labeling of oligonucleotide probes with europium chelates is presented. A modified deoxycytidine phosphoramidite is used to introduce multiple reactive amino groups to the oligonucleotide during the synthesis phase. Upon deprotection and purification of the modified oligonucleotide, an isothiocyanate derivative of a stable Eu chelate is reacted with the primary amino groups. The labeling technology enables the coupling of a high number of Eu chelates to a single probe. The melting temperatures and hybridization efficiencies of the oligonucleotides are not significantly altered by the labeling process. However, hybridization kinetics of the oligonucleotides are affected by the introduction of multiple modified deoxycytidine residues. In a solid-phase hybridization assay up to 10(7) target molecules can be detected.
authors.affiliation: Wallac Oy, Turku, Finland.
authors.forename: P
authors.initials: P
authors.lastname: Dahlén
authors.forename: L
authors.initials: L
authors.lastname: Liukkonen
authors.forename: M
authors.initials: M
authors.lastname: Kwiatkowski
authors.forename: P
authors.initials: P
authors.lastname: Hurskainen
authors.forename: A
authors.initials: A
authors.lastname: Iitiä
authors.forename: H
authors.initials: H
authors.lastname: Siitari
authors.forename: J
authors.initials: J
authors.lastname: Ylikoski
authors.forename: V M
authors.initials: VM
authors.lastname: Mukkala
authors.forename: T
authors.initials: T
authors.lastname: Lövgren
mesh_terms: D001483:Base Sequence; D002614:Chelating Agents; D005063:Europium; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000138:chemical synthesis* / Q000235:genetics; D000515:alpha 1-Antitrypsin / Q000235:genetics
chemical_list: D002614:Chelating Agents; D015345:Oligonucleotide Probes; D000515:alpha 1-Antitrypsin; D005063:Europium