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Updated: Jun 6, 2026
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Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
Published on: March 16, 2011
Selection of targeted mutants from a library of randomly mutagenized ES cells
Kyoji Horie1, George Gaitanaris, Alexander Gragerov
1Omeros Corporation, Seattle, WA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|November 17, 2010
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.
