Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4410

This Protocol
Right arrow Full Text
Right arrow Update/discuss this protocolDiscussion icon
Right arrow Alert me when this protocol is cited
Right arrow Alert me when comments are published
Right arrow Alert me if a correction is posted
Services
Right arrow Similar protocols in this database
Right arrow Alert me to new releases of protocols
Right arrow Save to Personal Folders
Right arrow Download to citation manager
Right arrow Printer-friendly versionPrinter-friendly version
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Nagy, A.
Right arrow Articles by Behringer, R.
Right arrow Search for Related Content
PubMed
Right arrow Articles by Nagy, A.
Right arrow Articles by Behringer, R.
Related Collections
Right arrow CSHL Molecular Embryology of the Mouse Course
Right arrow Laboratory Organisms, general
Right arrow Mouse
Right arrow Genotypic Analysis
Right arrow Developmental Biology
Right arrow Stem Cells, general
Right arrow Embryonic Stem Cells
Right arrowRelated Protocols
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?
BSN globe

protocolProtocol

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


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?

Related Protocols

Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomycin C
Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer
CSH Protocols 2006: 4399. [Extract] [Full Text]

Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with {gamma}-Irradiation
Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer
CSH Protocols 2006: 4400. [Extract] [Full Text]

Electroporating DNA into Embryonic Stem (ES) Cells and Selection Methods
Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer
CSH Protocols 2006: 4409. [Extract] [Full Text]

Freezing Embryonic Stem (ES) Cells in 96-Well Plates
Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer
CSH Protocols 2006: 4411. [Extract] [Full Text]