In the post-genomic era, PCR has become the method of choice not only for cloning existing genes, but also for generating a wide array of novel genes by mutagenesis and/or recombination within the genes of interest. PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long-distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination and to clone the challenging uncharacterized DNA flanking a known DNA fragment. Powerful applications of PCR in library construction and sublibrary generation and screening are also presented.
Authoritative and up-to-date, PCR Cloning Protocols, Second Edition, constitutes a gold-standard collection of the fastest, simplest, and most popular methods for isolating genes from all biological samples and creating novel genes from them by mutagenesis/recombination-essential methods for today's study of functional genomics, gene expression, protein structure-function relationships, protein engineering, and molecular evolution.