A simple and efficient method for site-directed mutagenesis with double-stranded plasmid DNA.
A general, simple and efficient method for preparing site-specific mutations in double-stranded plasmid DNA without the need for special plasmids, bacterial strains or reagents is described. Only one synthetic oligonucleotide for each mutation is required, subcloning is unnecessary and a high efficiency of mutation (58-97%) was obtained. If two synthetic oligonucleotide primers are used, two separate mutations can be simultaneously created in a single reaction tube.
Department of Molecular Genetics and Microbiology, UMDNJ-Robert Wood Johnson Medical School, Piscataway 08854-5635.
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D
Lai
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X
Zhu
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Pestka
D001483:Base Sequence; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids
D004269:DNA, Bacterial