Streamlined high-throughput cloning protocol to generate arrayed mutant libraries.
STAR Protoc
; 4(1): 101930, 2023 03 17.
Article
em En
| MEDLINE
| ID: mdl-36520626
ABSTRACT
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 sequence ready plasmid) in 3 days.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Reação em Cadeia da Polimerase
Idioma:
En
Revista:
STAR Protoc
Ano de publicação:
2023
Tipo de documento:
Article