Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4426

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Assembling Aggregates between Embryonic Stem (ES) Cells and Tetraploid Embryos

Andras Nagy, Marina Gertsenstein, Kristina Vintersten and Richard Behringer

This protocol was adapted from "Production of Chimeras," Chapter 11, 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 15% of the full text of this article appears below.


INTRODUCTION

This protocol describes a method for producing ES cell-tetraploid embryo chimeras. It requires the timed combination of four-cell-stage tetraploid embryo production and the procedure for ES cell-diploid embryo aggregation in which diploid embryos are replaced with tetraploid embryos. The resulting chimeras can be used to analyze the embryonic versus extraembryonic phenotype of a mutation.


MATERIALS

Reagents

Embryos, tetraploid (see Production of Tetraploid Embryos)

ES cells (see Preparing Embryonic Stem (ES) Cells for Aggregation)

Gelatin, 0.1% solution (e.g., Sigma) in H2O, autoclaved

recipe KSOM-AA mediumor other embryo culture medium

Equipment

. . . [Full Text of this Article]


METHOD


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