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Selection of Targeted Mutants from a Library of Randomly Mutagenized ES Cells

  • Kyoji Horie
  • George Gaitanaris
  • Alexander GragerovEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 693)

Abstract

A method for random relatively unbiased mutagenesis of ES cells with a mutagenic retroviral vector is described. An orderly assembly of mutant ES cells in multi-well plates is generated. 3D pooling of the wells of the assembly allows quick PCR search for insertions in genes of interest. Mutant ES cell clones are then isolated from the positive wells and used to produce mutant animals using conventional techniques.

Key words

Retroviral insertion Random mutagenesis ES cell Knock-out mice 3D pooling Gene targeting 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Kyoji Horie
    • 1
  • George Gaitanaris
    • 1
  • Alexander Gragerov
    • 1
    Email author
  1. 1.Omeros CorporationSeattleUSA

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