Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4410

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Isolating Individual Embryonic Stem (ES) Cell Colonies by Picking

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

This protocol describes a method for isolating individual ES cell colonies by picking after they have grown on selective medium.


MATERIALS

Reagents

ES cells under selection after DNA electroporation (see Electroporating DNA into Embryonic Stem (ES) Cells and Selection Methods)

recipe ES cell culture medium (ES-DMEM)

Gelatin, 0.1% solution (e.g., Sigma) in tissue culture-grade H2O

caution G418 (e.g., Geneticin, Invitrogen; Sigma) (200 µg/ml final concentration of the active ingredient in ES-DMEM)

Mouse embryonic fibroblast (MEF) or neo-resistant STO feeder cells (see Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomycin C or Preparing Feeder Cell . . . [Full Text of this Article]

Equipment


METHOD


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