Multiple end labeling of oligonucleotides with terbium chelate-substituted psoralen for time-resolved fluorescence detection.
A new procedure for the photochemical functionalization and the subsequent nonradioactive labeling of synthetic oligonucleotides with psoralen derivatives was developed where a double-stranded poly(A-T) tail is attached to the 5'- or 3'-end of the oligonucleotide to be labeled. The double-stranded poly(A-T) tail is covalently crosslinked by psoralen molecules which carry reactive thiol or amino groups for the attachment of labels. A NH2-specific terbium chelate exhibiting long-lived fluorescence was attached to the functional groups of the intercalated psoralen molecules. Oligonucleotides substituted in this way hybridize readily and can be sensitively detected by time-resolved fluorescence measurements.
Oser
A
A
Arbeitsgruppe Zellbiochemie, Max-Planck-Institut für Biochemie, Martinsried, Federal Republic of Germany.
Collasius
M
M
Valet
G
G
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005363:Ficusin / Q000737:chemistry; D005453:Fluorescence; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000032:analysis* / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013725:Terbium / Q000737:chemistry; D013997:Time Factors
D009841:Oligonucleotides; D013725:Terbium; D005363:Ficusin