Transgenesis Techniques: 18

Introduction and Topical Reviews. Introduction to Transgenesis. Transgenic Rodents and the Study of the Central Nervous System. Transgenic Animals and the Study of Cancer. Transgenic Animals and the Study of the Immune System. The Commercial and Agricultural Applications of Animal Transgenesis. Transgenesis in Invertebrate and Lower Vertebrate Species. P Element-Mediated Germ-Line Transformation of Drosophila. Gene Transfer by Microinjection in the Zebra Fish Brachydanio rerio. Transgenic Induction in Salmonid and Tilapia Fish. Transgenesis in the Mouse. An Overview of Transgenic Mouse Production. Transgene Design. Isolation of DNA Fragments for Microinjection. Establishing a Colony for Efficient Production of Transgenic Mice. Mating Mice. Anesthetizing Mice. Vasectomizing a Mouse. Preparation of Culture Media for Fertilized One-Cell Mouse Eggs. Collection of Fertilized One-Cell Mouse Eggs for Microinjection. Microinjection of Cloned DNA Fragments into Fertilized One-Cell Mouse Eggs: Manual Injection; Automatic Injection. Delivery of Microinjected Eggs to Surrogate Mothers by Oviduct Transfer. Caesarean Section and Fostering. In Vitro Fertilization of Mouse Eggs. The In Vitro Isolation of Murine Embryonic Stem Cells. Production of Chimeras Derived from Murine Embryonic Stem Cells. Cryopreservation of Transgenic Mouse Lines. Transgenesis in the Rat. Introduction to the Physiology and Husbandry of the Rat. Mating of Rats. Vasectomy of the Male Rat. Harvest of Fertilized One-Cell Rat Eggs. Implantation of Microinjected Eggs. Cryopreservation of Transgenic Rat Lines. Transgenesis in a Domestic Species-The Sheep. Production of Transgenic Sheep. Cryopreservation of Transgenic Sheep Lines. Genomic Analysis of Transgenic Animals. Isolation of Genomic DNA from Tail Tissue. Slot Blotting of Genomic Tail DNA. Genomic Analysis of Transgenic Animals by the Polymerase Chain Reaction. Genomic Analysis of Transgenic Animals: Southern Blotting. Expression Analysis of Transgenic Animals. The Preparation of RNA from the Tissues of Transgenic Animals. Northern Blotting. RNA Slot Blotting. Analysis of Gene Expression by PCR. Nuclear Run-On Analysis of Transcription. S1 Nuclease Protection Mapping. Primer Extension. In Situ Hybridization Analysis of Transgenic Tissues. Immunohistochemical Analysis of Transgene Expression. Postmortem Examination of Transgenic Mice. Histological Examination of Transgenic Mice. Reporter Enzyme Assays. Appendix. Restriction Endonuclease Digestion of DNA. Agarose Gel Electrophoresis. Working with Radioactivity in the Transgenic Animal Laboratory. Random Primed Labeling of DNA. Filter Hybridization. DNA Transfection. Index.