Fast one-step procedure for the detection of nucleic acids in situ by primer-induced sequence-specific labeling with fluorescein-12-dUTP.
We provide fast, simple, one-step procedures for sequence-specific detection of nucleic acids in situ. Tandem repeat sequences in DNA are stained within 30 min, and mRNA is stained within 2 h. The procedures are based on the incorporation of the newly available fluorescein-labeled dUTP into DNA synthesized in situ by primed in situ labeling, with denatured fragments of cloned DNA or oligonucleotides as primers. The extreme speed and simplicity of the reaction make it attractive for automatization in routine laboratory procedures and opens up new diagnostic possibilities.
Koch
J
J
Cytometry Division, University of Aarhus, Denmark.
Mogensen
J
J
Pedersen
S
S
Fischer
H
H
Hindkjaer
J
J
Kølvraa
S
S
Bolund
L
L
D004247:DNA / Q000032:analysis*; D015342:DNA Probes / Q000378:metabolism*; D003856:Deoxyuracil Nucleotides / Q000378:metabolism*; D005452:Fluoresceins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009841:Oligonucleotides / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis*; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics
D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D005452:Fluoresceins; D009841:Oligonucleotides; D012333:RNA, Messenger; C069632:fluorescein-11-deoxyuridine triphosphate; D004247:DNA