Plasmid-based one-pot saturation mutagenesis

Emily E. Wrenbeck; Justin R. Klesmith; James A. Stapleton; Adebola Adeniran; Keith E.J. Tyo; Timothy A. Whitehead

doi:10.1038/nmeth.4029

Plasmid-based one-pot saturation mutagenesis

Emily E. Wrenbeck, Justin R. Klesmith, James A. Stapleton, Adebola Adeniran, Keith E.J. Tyo, Timothy A. Whitehead

Chemical Engineering and Materials Science

Research output: Contribution to journal › Article › peer-review

118 Scopus citations

Abstract

Deep mutational scanning is a foundational tool for addressing the functional consequences of large numbers of mutants, but a more efficient and accessible method for construction of user-defined mutagenesis libraries is needed. Here we present nicking mutagenesis, a robust, single-day, one-pot saturation mutagenesis method performed on routinely prepped plasmid dsDNA. The method can be used to produce comprehensive or single- or multi-site saturation mutagenesis libraries.

Original language	English (US)
Pages (from-to)	928-930
Number of pages	3
Journal	Nature Methods
Volume	13
Issue number	11
DOIs	https://doi.org/10.1038/nmeth.4029
State	Published - Nov 1 2016

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© 2016 Nature America, Inc., part odf Springer Nature. All rights reserved.

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http://europepmc.org/articles/pmc5666567

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Plasmid-based one-pot saturation mutagenesis

Abstract

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