Laboratory Protocols for Conditional Gene Targeting
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Laboratory Protocols for Conditional Gene Targeting

Laboratory Protocols for Conditional Gene Targeting


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About the Book

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 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;

Table of Contents:
Section 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 reading

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


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Product Details
  • ISBN-13: 9780199636778
  • Publisher: Oxford University Press
  • Publisher Imprint: Oxford University Press
  • Height: 230 mm
  • No of Pages: 180
  • Weight: 554 gr
  • ISBN-10: 019963677X
  • Publisher Date: 03 Jul 1997
  • Binding: Hardback
  • Language: English
  • Spine Width: 18 mm
  • Width: 192 mm


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