The generation of a single nick per plasmid molecule using restriction endonucleases with multiple recognition sites.
Some restriction endonucleases generate a single-stranded nick at their recognition sequences in the presence of ethidium bromide (EtBr). This nick can then be extended to a single-stranded gap in which mutations can be introduced by a variety of techniques. To date, the templates used in these studies have largely contained a single recognition site for a given enzyme. Therefore, we have extended these studies to twelve enzymes for which multiple recognition sites exist in the template and show that, under appropriate conditions, one single-stranded nick is introduced per plasmid molecule.
D000818:Animals; D001483:Base Sequence; D004247:DNA; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004587:Electrophoresis, Agar Gel; D005796:Genes; D010957:Plasmids; D013379:Substrate Specificity; D013698:Templates, Genetic; D014337:Trout
D004247:DNA; D004262:DNA Restriction Enzymes