This laboratory manual, published in cooperation with the International Society for Transgenic Technology (ISTT), provides almost all current methods that can be applied to the creation and analysis of genetically modified animals. The chapters have been contributed by leading scientists who are actively using the technology in their laboratories. Based on their first-hand experience the authors also provide helpful notes and troubleshooting sections. Topics range from standard techniques, such as pronuclear microinjection of DNA, to more sophisticated and modern methods, such as the derivation and establishment of embryonic stem (ES) cell lines, with defined inhibitors in cell culture medium. In addition, related topics with relevance to the field are addressed, including global web-based resources, legal issues, colony management, shipment of mice and embryos, and the three R's: refinement, reduction and replacement.

<p>Karen S. Canady: Patent and licensing issues in transgenic technology

Lluís Montoliu: Global Resources: Including Gene Trapped ES Cell Clones: Is Your Gene Already Knocked Out?
Eduardo Moltó, Cristina Vicente-García and Lluís Montoliu: Designing Transgenes for Optimal Expression
Thomas L. Saunders: Gene Targeting Vector Design for Embryonic Stem Cell Modifications
Thomas J. Fielder and Lluis Montoliu: Transgenic Production Benchmarks
Katja Becker and Boris Jerchow: Generation of Transgenic Mice by Pronuclear Microinjection
Séverine Ménoret, Séverine Remy, et al : Generation of Transgenic Rats using Microinjection of Plasmid DNA or Lentiviral vectors
Almudena Fernández, Diego Muñoz and Lluís Montoliu: Generation of Transgenic Animals by Use of YACs
Michael G. Zeidler, Margaret L. Van Keuren, and Thomas L. Saunders: BAC Transgenes, DNA Purification, and Transgenic Mouse Production
Carlos Lois: Generation of Transgenic Animals with Lentiviral Vectors
Aron Geurts, Lajos Mates, Darius Balciunas: Vertebrate Transgenesis by Transposition
Karen M. Chapman et al.: Rat Spermatogonial Stem Cell Mediated Gene Transfer
Sayaka Wakayama, Nguyen Van Thuan andTeruhiko Wakayama: Mouse Cloning by Nuclear Transfer
Elizabeth D. Hughes and Thomas L. Saunders: Gene Targeting in Embryonic Stem Cells
Wojtek Auerbach and Anna B. Auerbach: The Importance of Mouse ES Cell Line Selection
Marina Gertsenstein: Tetraploid Complementation Assay
Elizabeth Williams et al.: Combining ES cells with Embryos
Kristina Nagy and Jennifer Nichols: Derivation of Murine ES Cell Lines
Ping Li et al.: Rat Embryonic Stem Cell Derivation and Propagation
Han Li et al.: Induced pluripotency: generation of iPS cells from mouse embryonic fibroblasts
Anna B. Auerbach, Peter J. Romanienko, and Willie H. Mark: The Preparation and Analysis of DNA for Use in Transgenic Technology
Karen Brennan: Colony Management
B. Pintado and J Hourcade: Cryopreservation
ShirleyPease: Shipment of Mice and Embryos
J.M. Sztein, R.J. Kastenmayer, K.A. Perdue: Pathogen Free Mouse Rederivation by IVF, Natural Mating and Hysterectomy
Jan Parker-Thornburg: Refinement, Reduction and Replacement.</p>