Rapid and Reliable Site-Directed Mutagenesis Using Kunkel's Approach -- Site-Directed Mutagenesis Using Altered /gb-Lactamase Specificity -- Site-Directed Mutagenesis Facilitated by DpnI Selection on Hemimethylated DNA -- Multiple Site-Directed Mutagenesis In Vitro -- Two-Stage Polymerase Chain Reaction Protocol Allowing Introduction of Multiple Mutations, Deletions, and Insertions, Using QuikChangeTM Site-Directed Mutagenesis -- Efficient and Accurate Site-Directed Mutagenesis of Large Plasmids -- Combining Site-Specific Chemical Modification with Site-Directed Mutagenesis -- Site-Directed Mutagenesis Mediated by a Single Polymerase Chain Reaction Product -- Megaprimer Method for Polymerase Chain Reaction-Mediated Generation of Specific Mutations in DNA -- Generation of Epitope-Tagged Proteins by Inverse Polymerase Chain Reaction Mutagenesis -- Site-Directed Mutagenesis by Polymerase Chain Reaction -- Generation of Multiple Site-Specific Mutations by Polymerase Chain Reaction -- Phenotypic Expression of Polymerase Chain Reaction-Generated Random Mutations in a Foreign Gene After its Introduction into an Acinetobacter Chromosome by Natural Transformation -- Polymerase Chain Reaction-Mediated Mutagenesis in Sequences Resistant to Homogeneous Amplification -- Polymerase Chain Reaction-Based Signature-Tagged Mutagenesis -- High-Throughput Scanning Mutagenesis by Recombination Polymerase Chain Reaction -- In Vitro Scanning-Saturation Mutagenesis -- Random Transposon Mutagenesis of Large DNA Molecules in Escherichia coli -- Random Chromosomal Gene Disruption Using Cassette Mutagenesis -- Transplacement Mutagenesis -- Preparation of Transposon Insertion Lines and Determination of Insertion Sites in Arabidopsis Genome -- Evolutionary Molecular Engineering by Random Elongation Mutagenesis -- Random Mutagenesis for Protein Breeding -- DNA Shuffling and Family Shuffling for In Vitro Gene Evolution -- Mutagenic Polymerase Chain Reaction of Protein-Coding Genes for In Vitro Evolution.

In the post-genome era, in vitro mutagenesis has emerged as the critically important tool used by molecular biologists in establishing the functions of components of the proteome. In this second edition of In Vitro Mutagenesis Protocols, active researchers with proven track records describe in stepwise fashion their advanced mutagenesis techniques. Each contributor focuses on improvements to conventional site-directed mutagenesis, with chapters being devoted to chemical site-directed mutagenesis; PCR-based mutagenesis and the modifications that allow high-throughput experiments; and mutagenesis based on gene disruption that is both in vitro- and in situ-based. Additional methods are provided for in vitro gene evolution; for gene disruption based on transposon, recombination, and cassette mutagenesis; and for facilitating the introduction of multiple mutations. Each readily reproducible technique includes detailed step-by-step instructions, tips on pitfalls to avoid, and notes on reagents and suppliers. Time-tested and highly practical, the techniques in In Vitro Mutagenesis Protocols, Second Edition offer today's molecular biologists a rich compendium of reliable and powerful techniques with which to illuminate the proteome.