Xenopus Protocols

The 2nd edition of this Methods in Molecular Biology(TM) volume provides novel approaches inspired by Xenopus tropicalis and the sequencing of its genome. Offers a toolkit enabling laboratories to maximize the power of this extraordinary experimental system.

During the 20 th century Xenopus has flourished as an accessible experimental model system for studying vertebrate biology. The second edition of Xenopus Protocols:Post-Genomic Approaches expands upon the previous edition with novel approaches inspired by Xenopus tropicalis and the sequencing of its genome . Xenopus tropicalis genomics and tractable genetics combined with the unique experimental advantages of Xenopus laevis and their distinctive phylogenetic position make them unique model systems for vertebrate embryonic development in the 21 st century. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Xenopus Protocols: Post-Genomic Approaches, Second Edition provides a toolkit that will enable every laboratory to maximize the power of this extraordinary experimental system in the post-genomic age.

Focuses on post-genomic approaches to the model system of Xenopus Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

Xenopus tropicalis as a Model Organism for Genetics and Genomics: Past, Present and Future.-Husbandry of Xenopus tropicalis .-Generating Diploid Embryos from Xenopus tropicalis .-Navigating the Xenopus tropicalis Genome.-Genetic Analysis of Xenopus tropicalis .-Forward Genetic Screens in Xenopus using Transposon-mediated Insertional Mutagenesis.-Targeted Gene Disruption with Engineered Zinc-finger Nucleases (ZFNs).-Reverse Genetic Studies using Antisense Morpholino Oligonucleotides.-Chemical Genetics and Drug Discovery in Xenopus.-Maternal mRNA Knock-down Studies: Antisense Experiments using the Host-transfer Technique in Xenopus laevis and Xenopus tropicalis .-Generating Transgenic Frog Embryos by Restriction Enzyme Mediated Integration (REMI).-A Simple Method of Transgenesis using I- Sce I meganuclease in Xenopus .-Using FC31 Integrase to Mediate Insertion of DNA in Xenopus Embryos.- Xenopus Transgenics: Methods using Transposons.-Comparative Genomics-based Identification and Analysis of Cis-regulatory Elements.-Tet-On Binary Systems for Tissue-specific and Inducible Transgene Expression.-Chromatin Immunoprecipitation Analysis of Xenopus Embryos.-Transcriptomics Using Next Generation Sequencing Technologies.-Databases of Gene Expression in Xenopus Development.-Investigating Alternative RNA Splicing in Xenopus .-Immunoisolation of Protein Complexes from Xenopus .-Complementary Proteomic Analysis of Protein Complexes.-Antibody Development and use in Chromogenic and Fluorescent Immunostaining.-Multiple fluorescent in situ mRNA Hybridization (FISH) on Whole Mounts and Sections.-Methods to Analyze microRNA Expression and Function During Xenopus Development.-A Bromodeoxyuridine (BrdU) Based Protocol for Characterizing Proliferating Progenitors in Xenopus Embryos.-Microscopy Tools for Quantifying Developmental Dynamics in XenopusEmbryos.-Mathematical Modelling of Gene RegulatoryNetworks in Xenopus Development.-Stem-cell-like Embryonic Explants to Study Cardiac Development.-Studying Regeneration in Xenopus .-On-line Resources for Xenopus. Mathematical Modelling of Gene RegulatoryNetworks in Xenopus Development.-Stem-cell-like Embryonic Explants to Study Cardiac Development.-Studying Regeneration in Xenopus .-On-line Resources for Xenopus.