With the completion of the mouse and human genome sequences, the functional characterization of every mammalian gene and the deciphering of their molecular interaction network has become the next major challenge, and the mouse genome provides a convenient and vital model for these studies. In "Gene Knockout Protocols, Second Edition", distinguished contributors with extensive experience in the gene targeting and mouse genetics fields reveal a comprehensive collection of step-by-step laboratory protocols. Emphasizing the many new mutagenesis techniques developed over the last seven years, the volume covers topics such as gene modification in ES cells, stem cell manipulation, the generation of genetically engineered mice, and mutant phenotype analysis. Written in the highly successful Methods in Molecular Biologya series format, chapters contain brief introductions to their respective subjects, lists of the necessary materials and reagents, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Fully updated and authoritative, "Gene Knockout Protocols, Second Edition" provides a unique collection of expert bench protocols certain to be a valuable resource for those just entering the field of gene targeting but also for all genetic researchers today.