Article
Site-directed mutant libraries for isolating minimal mutations yielding functional changes.
Protein engineering, design & selection : PEDS - 1 May 2017
Chung Dong Hee, Potter Sarah C, Tanomrat Ammon C, Ravikumar Krishnakumar M, Toney Michael D
Abstract excerpt
Powerful, facile new ways to create libraries of site-directed mutants are demonstrated. These include: (1) one-pot-PCR, (2) multi-pot-PCR, and (3) split-mix-PCR. One-pot-PCR uses mutant oligonucleotides to generate megaprimers in situ, and it was used to randomly incorporate 28 mutations in a gabT gene in a single reaction. In more difficult cases, multi-pot-PCR can be employed: mutant megaprimers are...
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