FastTag Nucleic Acid Labeling System: a versatile method for incorporating haptens, fluorochromes and affinity ligands into DNA, RNA and oligonucleotides.
The FastTag Nucleic Acid Labeling System couples haptens, fluorochromes or affinity ligands to any nucleic acid by attaching a universal, photo-or heat-activatable moiety to which any sulfhydryl-reactive compound can be linked. To demonstrate the versatility of the FastTag system, we have labeled DNA, RNA and oligonucleotide probes with a variety of maleimide-coupled moieties and have used these probes in several 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 detected the corresponding sequences in human chromosome spreads and interphase nuclei by fluorescence in situ hybridization. An antisense oligonucleotide probe cocktail complementary to human proinsulin transcripts labeled using FastTag DNP (dinitrophenyl) detected, in situ, the appropriate transcripts in pancreatic tissue sections. Oligonucleotide primers labeled with FastTag FL were used to PCR-amplify a genomic DNA fragment, which was then detected immunologically. Finally, we discuss how DNA labeled with FastTag Fucose can be bound to a fucose-binding affinity matrix and eluted under mild conditions.
Daniel
S G
SG
Vector Laboratories, Inc., Burlingame, CA, USA.
Westling
M E
ME
Moss
M S
MS
Kanagy
B D
BD
D000345:Affinity Labels / Q000138:chemical synthesis / Q000378:metabolism*; D001386:Azides / Q000138:chemical synthesis / Q000378:metabolism; D015139:Blotting, Southern; D015342:DNA Probes / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000378:metabolism*; D006241:Haptens / Q000378:metabolism*; D006801:Humans; D017403:In Situ Hybridization; D008024:Ligands; D015345:Oligonucleotide Probes / Q000378:metabolism; D016133:Polymerase Chain Reaction; D015347:RNA Probes / Q000378:metabolism*
D000345:Affinity Labels; D001386:Azides; D015342:DNA Probes; D005456:Fluorescent Dyes; D006241:Haptens; D008024:Ligands; D015345:Oligonucleotide Probes; D015347:RNA Probes