Zebrafish, 2009 Methods and Protocols Methods in Molecular Biology Series, Vol. 546

Langue : Anglais

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In the last 20 years, research activity using the zebrafish Danio rerio has increased dramatically, due in part to the ease of breeding and raising them, their genetic tractability, embryonic accessibility, and their imaging potential. In Zebrafish: Methods and Protocols, expert researchers provide a collection of experimental techniques for the genetic modification, manipulation, and in vivo live imaging of zebrafish. This volume collects sophisticated and practical techniques, including several not yet widely known, from laboratories around the world. Written in the highly successful Methods in Molecular Biology? series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and accessible, Zebrafish: Methods and Protocols seeks to aid and inspire scientists utilizing this versatile animal model in their research.

Mutagenesis and Mutants.- Highly Efficient ENU Mutagenesis in Zebrafish.- Production of Pseudotyped Retrovirus and the Generation of Proviral Transgenic Zebrafish.- Undertaking a Successful Gynogenetic Haploid Screen in Zebrafish.- Cryopreservation and In Vitro Fertilization at the Zebrafish International Resource Center.- Transgenesis.- Transient and Stable Transgenesis Using Tol2 Transposon Vectors.- Analysis of Genes and Genome by the Tol2-Mediated Gene and Enhancer Trap Methods.- Bacterial Artificial Chromosome Transgenesis for Zebrafish.- Simple and Efficient Transgenesis with Meganuclease Constructs in Zebrafish.- Tissue-Specific Manipulations.- Nitroreductase-Mediated Cell Ablation in Transgenic Zebrafish Embryos.- Focal Electroporation in Zebrafish Embryos and Larvae.- Tissue Micromanipulation in Zebrafish Embryos.- Analyzing Gene Expression.- Zebrafish Spotted-Microarray for Genome-Wide Expression Profiling Experiments. Part I: Array Printing and Hybridization.- Zebrafish Spotted-Microarray for Genome-Wide Expression Profiling Experiments: Data Acquisition and Analysis.- Validating microRNA Target Transcripts Using Zebrafish Assays.- Imaging.- Multiple Embryo Time-Lapse Imaging of Zebrafish Development.- Live Cell Imaging of Zebrafish Leukocytes.- Imaging Zebrafish Embryos by Two-Photon Excitation Time-Lapse Microscopy.- Morphological Analysis of the Zebrafish Digestive System.- In Toto Imaging of Embryogenesis with Confocal Time-Lapse Microscopy.

Covers key techniques for zebrafish mutagenesis, transgenesis, and gene expression analysis and imaging Contains practical approaches to chemical and insertional mutagenesis, including frozen storage of zebrafish sperm Includes a range of transgenesis protocols (building transgenes from bacterial artificial chromosomes, transposon- and meganuclease-assisted transgenesis) Describes imaging techniques ranging from organ-specific examples to whole embryo in toto time-lapse imaging Includes supplementary material: sn.pub/extras