Transgenesis Techniques pp 185-197
Targeting Embryonic Stem Cells
Gene targeting of mouse embryonic stem cells facilitates the deliberate engineering of the mouse genome. The gene targeting vector is introduced into cells by electroporation, and cell clones carrying the targeting vector are obtained by drug selection. Clones can be screened effectively for correct recombination events by either Southern blot analysis with external and internal probes, or by long-range PCR. Positive clones are subsequently expanded and prepared for the generation of chimeric mice.
Key wordsGene Targeting ES cells Southern blot analysis Long-range PCR