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Stem Cells 2004;22:243-249 www.StemCells.com
© 2004 AlphaMed Press

EXPERIMENTAL PROTOCOLS FOR EMBRYONIC STEM CELL RESEARCH

Electroporation of Murine Embryonic Stem Cells: A Step-by-Step Guide

Dennie M. Tompers, Patricia A. Labosky

University of Pennsylvania, Department of Cell and Developmental Biology, Philadelphia, Pennsylvania, USA

Key Words. Embryonic stem cells • Mouse • Electroporation • Homologous recombination

Patricia A. Labosky, Ph.D., University of Pennsylvania, Dept. of Cell and Developmental Biology, 1109 BRBII/III, 421 Curie Boulevard, Philadelphia, Pennsylvania 19104-6058, USA. Telephone: 215-573-7547; Fax: 215-898-9871; e-mail: plabosky{at}mail.med.upenn.edu

ABSTRACT

The manipulation of embryonic stem (ES) cells to generate targeted mutations via homologous recombination has proved an invaluable resource for researchers from fields as diverse as embryology, immunology, physiology, and biophysics. The derivation and culture of these cells has been described in many places, but there is no substitute for hands-on experience. In lieu of that, here we provide a step-by-step guide for the technique of electroporation, complete with micrographs of the cells at each step of the process. This is intended to allow the first time ES cell worker, as well as the more experienced researcher, to view the morphology of ES cells under ideal conditions as well as when the cells are suboptimal and should be discarded.




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