User-defined single pot mutagenesis using unamplified oligo pools.
Protein Eng Des Sel
; 32(1): 41-45, 2019 09 10.
Article
em En
| MEDLINE
| ID: mdl-31297523
ABSTRACT
User-defined mutagenic libraries are fundamental for applied protein engineering workflows. Here we show that unamplified oligo pools can be used to prepare site saturation mutagenesis libraries from plasmid DNA with near-complete coverage of desired mutations and few off-target mutations. We find that oligo pools yield higher quality libraries when compared to individually synthesized degenerate oligos. We also show that multiple libraries can be multiplexed into a single oligo pool, making preparation of multiple libraries less expensive and more convenient. We provide software for automatic oligo pool design that can generate mutagenic oligos for saturating or focused libraries.
Palavras-chave
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Oligodesoxirribonucleotídeos
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Engenharia de Proteínas
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Biblioteca Gênica
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Mutagênese Sítio-Dirigida
Idioma:
En
Revista:
Protein Eng Des Sel
Assunto da revista:
BIOQUIMICA
/
BIOTECNOLOGIA
Ano de publicação:
2019
Tipo de documento:
Article
País de afiliação:
Estados Unidos