Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4409

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Electroporating DNA into Embryonic Stem (ES) Cells and Selection Methods

Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer

This protocol was adapted from "Introduction of Foreign DNA into Embryonic Stem Cells," Chapter 10, in Manipulating the Mouse Embryo, 3rd edition, by Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

The first 100 words of the full text of this article appear below.


INTRODUCTION

Electroporation is one of the most efficient methods for introducing DNA into ES cells. This protocol describes a method for electroporating DNA into ES cells, as well as selection methods. Pilot studies should be performed to optimize the conditions for each DNA construct. The selection method described here is one of the most complex. It involves targeting constructs in which the bacterial neomycin-resistance gene disrupts the coding sequence of the mouse gene.


MATERIALS

Reagents

DNA, Linear, 25 µg electroporation (circular DNA for transient expression of a transgene, e.g., Cre recombinase)

Electroporation buffer (Specialty Media) (optional)

ES cells (exponentially growing) (see Passage of . . . [Full Text of this Article]

Equipment


METHOD


TROUBLESHOOTING


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