Primer-directed mutagenesis of linearized plasmids.
Two novel mutagenesis techniques to specifically alter the sequence of plasmid DNA have been developed. In contrast to other primer-directed mutagenesis methods which require a single-stranded, closed-circular template, a linearized single strand was used as the mutagenesis template. The template is prepared by restriction enzyme digestion of covalently-closed-circular plasmid DNA. These methods are simple, require small amounts of plasmid DNA, and can result in a relatively high frequency of mutagenesis.
D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA; D004269:DNA, Bacterial / Q000235:genetics; D004277:DNA, Single-Stranded / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D009154:Mutation; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000138:chemical synthesis; D010957:Plasmids; D013698:Templates, Genetic; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D009841:Oligonucleotides; D004247:DNA