FASTTAG(TM) NUCLEIC-ACID LABELING SYSTEM - A VERSATILE METHOD FOR INCORPORATING HAPTENS, FLUOROCHROMES AND AFFINITY LIGANDS INTO DNA, RNA AND OLIGONUCLEOTIDES

The FastTag(TM) Nucleic Acid Labeling System couples haptens, fluoroch romes or affinity ligands to any nucleic acid by attaching a universal , photo- or heat-activatable moiety to which any sulfhydryl-reactive c ompound can be linked. To demonstrate the versatility of the FastTag s ystem, we have labeled DNA, RNA and oligonucleotide probes with a vari ety of maleimide-coupled moieties and have used these probes in severa l applications. in Southern hybridization analyses, RNA probes labeled using FastTag FL (fluorescein) detected 0.04 pg of target DNA. Human satellite DNA clones labeled using FastTag FL or FastTag Biotin detect ed the corresponding sequences in human chromosome spreads and interph ase nuclei by fluorescence in situ hybridization. An antisense oligonu cleotide probe cocktail complementary to human proinsulin transcripts labeled using FastTag DNP (dinitrophenyl) detected, in situ, the appro priate transcripts in pancreatic tissue sections. Oligonucleotide prim ers labeled with FastTag FL were used to PCR-amplify a genomic DNA fra gment, which was then detected immunologically. Finally, we discuss ho w DNA labeled with FastTag Fucose can be bound to a fucose-binding aff inity matrix and eluted under mild conditions.