Laboratory Protocols for Conditional Gene Targeting

Hardcover | June 1, 1997

byRaul M. Torres, Ralf KuhnForeword byKlaus Rajewsky

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Conditional 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 fieldas 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 selectionmarker 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 cellculture 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 needlesand holders; blastocyst preparation/injection; embryo aggregation; blastocyst transfer;

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From the Publisher

Conditional 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 fieldas well as more experienced researchers. Top...

Raul Torres is at Basel Institute for Immunology. Ralf Kuhn is at University of Cologne.

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Format:HardcoverDimensions:180 pages, 8.74 × 6.89 × 0.71 inPublished:June 1, 1997Publisher:Oxford University Press

The following ISBNs are associated with this title:

ISBN - 10:019963677X

ISBN - 13:9780199636778

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Table of Contents

Section 1: The Cre-loxP recombination system and gene targeting in ES cells1. Gene targeting strategies2. Cre-loxP recombination system3. Gene targeting vectors - consideration and use of loxP4. Nonselectablae modifications and removal of selection marker genes5. Large deletions6. Gene replacement7. Targeted integrations9. Conditional gene modification10. LoxP-containing transgenes11. Conditional gene targetingSection 2: Working with ES cells12. ES cell transfection overview13. Embryonic feeder cells14. ES cells - handling and use15. ES cell culture and transfection16. ES clone picking17. Identification of homologous recombinants18. Freezing ES clones19. Thawing ES clones20. Genomic DNA preparation21. Cre-mediated neomycin deletion in ES cells22. Generation of mice - preparation23. Mouse breeding for ES cell injection or aggregation24. Injection needles and holders25. Blastocyst preparation/injection26. Embryo aggregation27. Blastocyst transferAppendicesReferencesSelected references for further reading

Editorial Reviews

`With this book, the authors present the latest addition to the technical improvements in gene targeting.'BPI, February/March 1998