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Transgenic animal technologies and the ability to introduce functional genes into animals have revolutionized our ability to address complex biomedical and biological questions. This well-illustrated handbook covers the technical aspects of gene transfer - from molecular methods to whole animal considerations - for important laboratory and domestic animal species. It describes methodologies as employed by leading laboratories and is a key resource for researchers, as well as a tool for training technicians and students. This second edition incorporates updates on a variety of genetic engine.

Front Cover; Transgenic Animal Technology; Copyright Page; Contents; List of Contributors; Preface; One: Overview; 1 Introduction to Transgenic Animal Technology; I. Introduction; II. Historical Background; III. Applications and Overview of Text; References; Two: Transgenic Animal Production Focusing on the Mouse Model; 2 DNA Microinjection, Embryo Handling, and Germplasm Preservation; I. Introduction; II. General Methods; A. DNA Preparation and Purification; 1. DNA Construct/Fragment Structure; 2. Factors that Influence Transgenic Mouse Production; 3. Preparation of DNA for Microinjection.

A. Construction and Isolation of DNAb. DNA Purification; c. Purification and Quantification of DNA; B. Superovulation, Egg Culture, and Harvest; 1. Mouse Strains for Microinjection and Transgenic Models; a. Strains; b. Genetic Variability and Transgene Expression; 2. Superovulation of Mice; a. Pregnant Mare Serum Gonadotropin and Human Chorionic Gonadotropin; b. Follicle Stimulating Hormone and Luteinizing Hormone Delivered with Osmotic Pumps; 3. Production of Eggs from Superovulated Females; a. Colony; b. Influence of Litter Number; 4. In Vitro Culture of Superovulated Eggs; a. Equipment.

B. Atmosphere for Culturing Eggsc. Glass Pipettes for Manipulating Eggs; d. Media; e. Water Sources; f. Culture Dishes; 5. Harvesting Superovulated Eggs; a. Mating; b. Oviduct Dissection; c. Release of Eggs from the Oviduct; d. Egg Development and Microinjection Timing; e. Efficiency of Egg Culture; C. Microinjection Needles and Slides; 1. Types of Glass; 2. Preparation of Glass; 3. Pipette Pullers; 4. Injection Pipette Shapes; 5. Beveling Hollow Capillary Microinjection Needles; 6. Microforge Shaping of Pipettes; a. Manufacturers; b. Heating Filament; c. Holding Pipette; d. Injection Pipette.

7. Microinjection Slide for Eggsa. Slide Designs; b. Microinjection Media; c. Arrangement of DNA and Media Drops; D. Microinjection Equipment; 1. Microscopes; 2. Micromanipulators; a. Micropipette Holders; b. Micromanipulator Adjustments; c. Antivibration Tables; 3. Microinjection Systems; a. General; b. Oil-Driven Injection Assembly; 4. Oil-Driven Holding Pipettes; 5. Electronic Delivery Systems; E. Microinjection Procedure; 1. Preparation of Injection and Holding Pipettes (Micrometer Syringe System); a. Holding Pipette; b. Injection Pipette; c. Aligning the Injection Needle.

D. Loading DNA into the Injection Needle2. Preparing Eggs for Microinjection; 3. Steps for Efficient Manipulation of Eggs During Microinjection; a. General; b. Micromanipulators; c. Microinjection Needle; d. Controls and Egg Manipulation; 4. Visualizing and Injecting the Egg Pronuclei; a. Orientation; b. Egg Membrane Penetration with the Injection Needle; c. Filling the Pronuclei with DNA Solution; 5. Criteria for Microinjection to Yield Transgenic Mice; a. Treatment After Microinjection; b. Egg Lysis; c. Record Keeping; d. In Vitro Culture of Microinjected Eggs; F. Egg Transfer.

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