Mutagenesis method
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United States of America Patent
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Sep 27, 2016
Grant Date -
Feb 21, 2013
app pub date -
Dec 21, 2010
filing date -
Dec 21, 2009
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Abstract
The present invention provides a mutagenesis method wherein a nucleic acid molecule is mutagenized with at least one mutagenesis primer in a primer extension reaction and subsequently amplified by rolling circle amplification (RCA). The method involves the step of rendering the template strand unfavorable for RCA. The method involves steps leading to selective amplification of only the mutated strand by a strand-displacing DNA polymerase. Multiple copies of the mutated plasmids are generated during multiple-primed RCA and the resulting DNA is transformed for use. The method is suitable for mutating both single-stranded and double-stranded DNA. The present invention also provides a kit for use in the mutagenesis method.
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