Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4418

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Injecting Blastocysts

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

This protocol describes a method for injecting mouse blastocysts with embryonic stem (ES) cells to produce chimeras.


MATERIALS

Reagents

Blastocyst-stage embryos (see Collecting Blastocysts)

ES cells prepared according to Preparation of Embryonic Stem (ES) Cells for Injection

recipe M16

or

recipe KSOM-AA medium

Equipment

Holding pipettes (see Making Holding Pipettes)

Humidified incubator, at 37°C, 5% CO2, 95% air

Injection chamber (see Microinjection Setup)

Injection pipette (see Making Injection Pipettes)

Microscope/micromanipulator setup (see Microinjection Setup)

Mineral oil, embryo-tested light

Transfer pipettes and mouth pipette device (see Making Pipettes from Hard Glass Capillaries)


METHOD

1. Precool the injection chamber on the . . . [Full Text of this Article]


TROUBLESHOOTING


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