Controlled partial restriction digestions of DNA by competition with modification methyltransferases.
Competitive reactions, using defined ratios of DNA restriction methyltransferase to endonuclease, are shown to result in reliable partial restriction digests of DNA. This method is suitable over a wide range of DNA concentrations and works on DNA in liquid or embedded in agarose. Simultaneous methylase/endonuclease reactions using endonucleases that cleave human DNA very infrequently, such as ClaI or NotI, should generate very large discrete partial DNA fragments suitable for physical mapping in the million base-pair range. Another possible application of methylase/endonuclease competitive reactions is the production of defined partial digests for making cosmid, lambda, or other genomic libraries.
Hanish
J
J
Department of Biochemistry and Molecular Biology, University of Chicago, Illinois 60637.
McClelland
M
M
D001345:Autoradiography; D010582:Bacteriophage lambda / Q000235:genetics*; D001667:Binding, Competitive; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D008780:Methyltransferases
D004279:DNA, Viral; D008780:Methyltransferases; D004262:DNA Restriction Enzymes