Plasmid-based one-pot saturation mutagenesis

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

Research output: Contribution to journalArticlepeer-review

49 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 languageEnglish (US)
Pages (from-to)928-930
Number of pages3
JournalNature Methods
Volume13
Issue number11
DOIs
StatePublished - Nov 1 2016

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