Primer-directed mutagenesis of linearized plasmids |
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Authors: | Stanley M. Hollenberg Jiunu S. Lai Joachim L. Weickmann Takayasu Date |
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Affiliation: | International Genetic Engineering, Inc., 1701 Colorado Avenue, Santa Monica, California 90404 USA |
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Abstract: | 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. |
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