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OverviewConditional gene targeting is an emerging technique for introducing mutations into the genes of laboratory mice. This collection of protocols comes from one of the world's premier laboratories for gene targeting The Institute of Genetics in Cologne and is intended for newcomers to the field as well as more experienced researchers. Topics covered include: the Cre/loxP recombination system and gene targeting in ES cells; gene targeting strategies; Cre/loxP recombination system; Gene targeting vectors consideration and use of loxP; nonselectable modifications and removal of selection marker genes; large deletions; gene replacement; chromosomal translocations; targeted integrations; conditional gene modification; LoxP-containing transgenes; conditional gene targeting; working with ES cells; ES cell transfection overview; embryonic feeder cells; ES cells handling and use; ES cell culture and transfection; ES clone picking; identification of homologous recombinants; freezing ES clones; thawing ES clones; genomic DNA preparation; cre-mediated neomycin deletion in ES cells; generation of mice preparation; mouse breeding for ES cell injection or aggregation; injection needles and holders; blastocyst preparation/injection; embryo aggregation; blastocyst transfer; Full Product DetailsAuthor: Raul M. Torres (Basel Institute for Immunology) , Ralf Kühn (Institute for Genetics, University of Cologne) , Klaus RajewskyPublisher: Oxford University Press Imprint: Oxford University Press Dimensions: Width: 19.20cm , Height: 1.80cm , Length: 23.00cm Weight: 0.554kg ISBN: 9780199636778ISBN 10: 019963677 Pages: 180 Publication Date: 03 July 1997 Audience: Professional and scholarly , Professional & Vocational Format: Hardback Publisher's Status: Out of Print Availability: Out of stock  Table of ContentsSection 1: The Cre-loxP recombination system and gene targeting in ES cells 1: Gene targeting strategies 2: Cre-loxP recombination system 3: Gene targeting vectors - consideration and use of loxP 4: Nonselectablae modifications and removal of selection marker genes 5: Large deletions 6: Gene replacement 7: Targeted integrations 9: Conditional gene modification 10: LoxP-containing transgenes 11: Conditional gene targeting Section 2: Working with ES cells 12: ES cell transfection overview 13: Embryonic feeder cells 14: ES cells - handling and use 15: ES cell culture and transfection 16: ES clone picking 17: Identification of homologous recombinants 18: Freezing ES clones 19: Thawing ES clones 20: Genomic DNA preparation 21: Cre-mediated neomycin deletion in ES cells 22: Generation of mice - preparation 23: Mouse breeding for ES cell injection or aggregation 24: Injection needles and holders 25: Blastocyst preparation/injection 26: Embryo aggregation 27: Blastocyst transfer Appendices References Selected references for further readingReviews<br> This outstanding laboratory manual focuses on sophisticated, state-of-the-art Cre/loxP recombination strategies for generating site-specific modifications of the mouse genome. . . . In the first half of this manual, the editors discuss the theory behind the Cre/loxP recombination system, and then describe in detail the various experimental strategies for using Cre/loxP for manipulating the mouse genome. . . . These sections made for enjoyable reading, and they provide a comprehensive view of the many ways one can manipulate the mouse genome. The second half of the book is devoted to detailed laboratory protocols for actually carrying out the strategies described in the previous section. . . . The appendix contains recipes for all solutions and media, maps of gene targeting plasmids, and even a listing of equipment vendors. The attention to detail is excellent. Both new and experienced investigators should find this manual indispensable reading. --The Quarterly Review of Biology<p><br> This outstanding laboratory manual focuses on sophisticated, state-of-the-art Cre/loxP recombination strategies for generating site-specific modifications of the mouse genome. . . . In the first half of this manual, the editors discuss the theory behind the Cre/loxP recombination system, and then describe in detail the various experimental strategies for using Cre/loxP for manipulating the mouse genome. . . . These sections made for enjoyable reading, and they provide a comprehensive view of the many ways one can manipulate the mouse genome. The second half of the book is devoted to detailed laboratory protocols for actually carrying out the strategies described in the previous section. . . . The appendix contains recipes for all solutions and media, maps of gene targeting plasmids, and even a listing of equipment vendors. The attention to detail is excellent. Both new and experienced investigators should find this manual indispensable reading. --The Quarterly Review of Biology<br> Author InformationTab Content 6Author Website:Countries AvailableAll regions |
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