Streamlined high-throughput cloning protocol to generate arrayed mutant libraries.
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ABSTRACT:
SUBMITTER: Sun KT
PROVIDER: S-EPMC9758493 | biostudies-literature | 2022 Dec
REPOSITORIES: biostudies-literature
Publications
Streamlined high-throughput cloning protocol to generate arrayed mutant libraries.
Sun Kerry T KT Patel Tark S TS Kim Justin J Tang Helen S H HSH Eskandari-Sedighi Ghazaleh G Sureshkumar Haresh H Schieve Dean D Mok S A SA
STAR protocols 20221213 1
Large-scale, site-directed mutagenesis enables rapid characterization of the biochemical and biological properties of proteins. Here, we present a cost-effective and adaptable cloning pipeline to generate arrayed gene libraries for a construct of interest. We detail steps to use an open access web app to automate the design of mutagenesis primers optimized for our cloning protocols in a 96-well plate format. The protocol allows most molecular biology labs to clone 96 mutants (from PCR to sequenc ...[more]
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