期刊
PROTEIN ENGINEERING DESIGN & SELECTION
卷 32, 期 1, 页码 41-45出版社
OXFORD UNIV PRESS
DOI: 10.1093/protein/gzz013
关键词
deep mutational scanning; directed evolution; nicking mutagenesis; oligo pools; saturation mutagenesis
资金
- National Institute Of Allergy And Infectious Diseases of the National Institutes of Health [R01AI141452]
- DARPA APT
- NIH T32 Biotechnology Training Grant [T32-GM110523]
- Defense Advanced Research Projects Agency (DARPA)
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.
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