Facile Site-Directed Mutagenesis of Large Constructs Using Gibson Isothermal DNA Assembly.
Methods Mol Biol
; 1498: 359-366, 2017.
Article
em En
| MEDLINE
| ID: mdl-27709588
ABSTRACT
Site-directed mutagenesis is a commonly used molecular biology technique to manipulate biological sequences, and is especially useful for studying sequence determinants of enzyme function or designing proteins with improved activity. We describe a strategy using Gibson Isothermal DNA Assembly to perform site-directed mutagenesis on large (>~20 kbp) constructs that are outside the effective range of standard techniques such as QuikChange II (Agilent Technologies), but more reliable than traditional cloning using restriction enzymes and ligation.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Mutagênese
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
Estados Unidos