A rapid and simple procedure for the preparation of homogeneously labeled DNA probes.
This report describes a novel technique for the preparation of homogeneously labeled DNA probes. The procedure uses a heteroduplex intermediate, which is directly prepared from a double-stranded plasmid, and therefore requires neither sub-cloning of the fragment of interest into a specialized vector nor the use of a synthetic primer. The technical advantages of the procedure are demonstrated by preparation of a probe able to determine the precise location and efficiency of utilization of the polyadenylation site used during transcription of the rat preproinsulin II gene in a transfected avian cell line.
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D004720:Endonucleases; D005786:Gene Expression Regulation; D007328:Insulin; D010957:Plasmids; D011061:Poly A / Q000235:genetics*; D011384:Proinsulin / Q000235:genetics*; D011498:Protein Precursors / Q000235:genetics*; D012323:RNA Processing, Post-Transcriptional; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics; D015719:Single-Strand Specific DNA and RNA Endonucleases
D007328:Insulin; D011498:Protein Precursors; D011994:Recombinant Proteins; D011061:Poly A; C013737:preproinsulin; D011384:Proinsulin; D004720:Endonucleases; D015719:Single-Strand Specific DNA and RNA Endonucleases