Transgenesis techniques : principles and protocols

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 -- Commercial and agricultural applications of animal transgenesis -- 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 -- 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. 1, Manual injection -- Microinjection of cloned DNA fragments into fertilized one-cell mouse eggs. 2, Automatic injection -- Delivery of microinjected eggs to surrogate mothers by oviduct transfer -- Caesarean section and fostering -- In vitro fertilization of mouse eggs -- In vitro isolation of murine embryonic stem cells -- Production of chimeras derived from murine embryonic stem cells -- Cryopreservation of transgenic mouse lines -- 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 -- Production of transgenic sheep -- Cryopreservation of transgenic sheep lines -- 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 -- 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 -- 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