Protocol

Microinjection Setup

This protocol was adapted from "Production of Transgenic Mice," Chapter 7 of 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 the microinjection setup for injecting mouse zygotes to produce transgenic mice.

MATERIALS

Reagents

DNA solution, 1-2 ng/µl, prepared by standard methods

M2 culture medium

M16 culture medium

Paraffin oil, light (e.g., Merck)

Equipment

Antivibration table (e.g., Newport)

Control unit for holding pipette (e.g., Eppendorf, Narishige, Sutter), or Tygon tubing and a mouth pipette device

Holding pipettes (see Making Holding Pipettes)

Injection chamber: see Step 3

Depending on the type chosen, one of these items will be needed: plastic Petri dish with glass bottom (Willco), glass depression slide or metal frame attached to glass slide with high-vacuum grease (Dow Corning).

. . . [Full Text of this Article]

METHOD

TROUBLESHOOTING