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Full Description
Hands-on researchers with proven track records describe in stepwise fashion their advanced mutagenesis techniques. The contributors focus on improvements to conventional site-directed mutagenesis, including a chapter on chemical site-directed mutagenesis, PCR-based mutagenesis and the modifications that allow high throughput mutagenesis experiments, and mutagenesis based on gene disruption (both in vitro- and in situ-based). Additional methods are provided for in vitro gene evolution; for gene disruption based on recombination, transposon, and casette mutagenesis; and for facilitating the introduction of multiple mutations. Time-tested and highly practical, the protocols in "In Vitro Mutagenesis Protocols, 2nd Edition" offer today's molecular biologists reliable and powerful techniques with which to illuminate the proteome.
Contents
Rapid and Reliable Site-Directed Mutagenesis Using Kunkel's Approach Priya Handa, Swapna Thanedar, and Umesh Varshney Site-Directed Mutagenesis Using Altered b-Lactamase Specificity Christine A. Andrews and Scott A. Lesley Site-Directed Mutagenesis Facilitated by DpnI Selection on Hemimethylated DNA Fusheng Li and James I. Mullins Multiple Site-Directed Mutagenesis In Vitro Yang-Gyun Kim and Stefan Maas Two-Stage Polymerase Chain Reaction Protocol Allowing Introduction of Multiple Mutations, Deletions, and Insertions, Using QuikChangea Site-Directed Mutagenesis Wenyan Wang and Bruce A. Malcolm Efficient and Accurate Site-Directed Mutagenesis of Large Plasmids Susan A. Nadin-Davis Combining Site-Specific Chemical Modification with Site-Directed Mutagenesis: Versatile Strategy to Move Beyond Structural Limitations of 20 Natural Amino Acids Side Chains in Protein Engineering Grace DeSantis and J. Bryan Jones Site-Directed Mutagenesis Mediated by a Single Polymerase Chain Reaction Product Xueni Chen, Weimin Liu, Ileana Quinto, and Giuseppe Scala Megaprimer Method for Polymerase Chain Reaction-Mediated Generation of Specific Mutations in DNA Jesper Brons-Poulson, Jane Nohr, and Leif Kongskov Larsen Generation of Epitope-Tagged Proteins by Inverse Polymerase Chain Reaction Mutagenesis Lucio Gama and Gerda E. Breitwieser Site-Directed Mutagenesis by Polymerase Chain Reaction Albert Jeltsch and Thomas Lanio Generation of Multiple Site-Specific Mutations by Polymerase Chain Reaction Amom Ruhikanta Meetei and M. R. S. Rao Phenotypic Expression of Polymerase Chain Reaction-Generated Random Mutations in a Foreign Gene After its Introduction into an Acinetobacter Chromosome by Natural Transformation David M. Young, Ruben G. Kok, and L. Nicholas Ornston Polymerase Chain Reaction-Mediated Mutagenesis in Sequences Resistant to Homogeneous Amplification Ross N. Nazar, P. D. Abeyrathne, and Robert V. A. Intine Polymerase Chain Reaction-Based Signature-Tagged Mutagenesis Dario E. Lehoux and Roger C. Levesque High-Throughput Scanning Mutagenesis by Recombination Polymerase Chain Reaction Stefan Howorka and Hagan Bayley In Vitro Scanning-Saturation Mutagenesis Jennifer A. Maynard, Gang Chen, George Georgiou, and Brent L. Iverson Random Transposon Mutagenesis of Large DNA Molecules in Escherichia coli Wolfram Brune Random Chromosomal Gene Disruption Using Cassette Mutagenesis French A. Lewis, III and Brian Dougherty Transplacement Mutagenesis: A Recombination-Based In Situ Mutagenesis Protocol Knut Woltjen, M. W. Todd Unger, and Derrick E. Rancourt Preparation of Transposon Insertion Lines and Determination of Insertion Sites in Arabidopsis Genome Takuya Ito, Reiko Motohashi, and Kazuo Shinozaki Evolutionary Molecular Engineering by Random Elongation Mutagenesis Tomoaki Matsuura, Tetsuya Yomo, and Itaru Urabe Random Mutagenesis for Protein Breeding Chris Fenton, Hao Xu, Evamaria I. Petersen, Steffen B. Petersen, and M. Raafat El-Gewely DNA Shuffling and Family Shuffling for In Vitro Gene Evolution Miho Kikuchi and Shigeaki Harayama Mutagenic Polymerase Chain Reaction of Protein-Coding Genes for In Vitro Evolution Ichiro Matsumura and Andrew D. Ellington
