Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4422

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Production of 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 100 words of the full text of this article appear below.


INTRODUCTION

Recently, tetraploid mouse embryos have become an important component of both embryo-embryo and embryonic stem (ES) cell-embryo chimeras. This protocol describes the production of tetraploid embryos by electrofusion.


MATERIALS

Reagents

Embryos, two-cell, collected at 1.5 days post coitum (dpc): see Step 1

recipe KSOM-AA medium or other embryo culture medium

Mannitol, 0.3 M (Sigma)

recipe M2 or other HEPES-buffered medium

Equipment

CF-150B pulse generator (BLS)

Dissecting microscope(s)

Electrode-chamber with 250-µm gap (e.g., GSS-250, BLS)

GSS-500 and GSS-1000 electrode chambers with 500- or 1000-µm gap distance, respectively, may also be used, and are available from the same company.

Incubator, humidified, at 37°C, 5% CO2

Microdrop culture . . . [Full Text of this Article]


METHOD


TROUBLESHOOTING


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