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Plasmid-based one-pot saturation mutagenesis.

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.

SUBMITTER: Wrenbeck EE

PROVIDER: S-EPMC5666567 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature

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Plasmid-based one-pot saturation mutagenesis.

Wrenbeck Emily E EE Klesmith Justin R JR Stapleton James A JA Adeniran Adebola A Tyo Keith E J KE Whitehead Timothy A TA

Nature methods 20161010 11

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. ...[more]

PMID: 27723752

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