E-Book Overview
A comprehensive compendium of cutting-edge protocols for the generation of molecular diversity. Described in step-by-step detail to ensure experimental success, these protocols include readily reproducible methods for random mutagenesis of entire genes or segments of genes, for homologous and nonhomologus recombination, and for constructing in vivo libraries in bacteria and yeast. In addition to the various protocols for creating libraries, this volume also describes ways to analyze libraries, particularly those made by recombination. An accompanying volume, Directed Enzyme Evolution: Screening and Selection Methods (ISBN: 1-58829-286-X), is devoted entirely to selection and screening methods that can be applied to the directed evolution of enzymes. Copy for Both Volumes Directed Evolution Library Creation: Methods and Protocols and Directed Enzyme Evolution: Screening and Selection Methods constitute an extraordinary collection of all the key methods used today for directed evolution research. Described in step-by-step detail to ensure robust experimental results, these methods will enable both newcomers and more experienced investigators to design and implement directed evolution strategies for the engineering of novel proteins. The first volume describes methods for the creation of mutated DNA molecules, or DNA libraries, encoding variants of desired proteins. The second volume describes methods for screening DNA libraries to isolate mutant proteins that exhibit a specified function.
E-Book Content
Methods in Molecular Biology
TM
VOLUME 231
Directed Evolution Library Creation Methods and Protocols Edited by
Frances H. Arnold George Georgiou
Mutant Libraries via PCR
3
1 Generating Mutant Libraries Using Error-Prone PCR Patrick C. Cirino, Kimberly M. Mayer, and Daisuke Umeno 1. Introduction Directed evolution h