S1 nuclease transcript mapping using sequenase-derived single-stranded probes.
We report here a simple method for the production of high specific activity single-stranded DNA probes for S1 nuclease transcript mapping. Plasmid DNA is used as in vitro DNA templates for probe production by primer extension using Sequenase. This allows the production of long single-stranded probes without the need to subclone restriction fragments into M13 vectors. The mapping of the 5' ends of the phage P1 mod gene transcripts is shown as an example.
Sharrocks
A D
AD
Department of Molecular Biology and Biotechnology, University of Sheffield, UK.
Hornby
D P
DP
D001435:Bacteriophages / Q000235:genetics; D015342:DNA Probes; D004277:DNA, Single-Stranded; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D005814:Genes, Viral; D008780:Methyltransferases / Q000235:genetics; D009710:Nucleotide Mapping / Q000379:methods; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism*
D015342:DNA Probes; D004277:DNA, Single-Stranded; D008780:Methyltransferases; C034970:bacteriophage T7 induced DNA polymerase; D004259:DNA-Directed DNA Polymerase; D015719:Single-Strand Specific DNA and RNA Endonucleases