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This second edition volume expands on the previous edition by presenting updated protocols for several of the techniques described in the first edition of Drosophila: Methods and Protocols and current methods that cover recent breakthroughs in Drosophila research. The book begins with a description of FlyBase--a database of genes and genomes--followed by the presentation of systems for versatile gene expression in the fly. The first few chapters in this book detail gene knockdown and editing, including CRISPR-Cas9 and protein knockdown. The next few chapters are devoted to methods describing live imaging of different tissues and organs, followed by chapters on how to quantify image data and how to probe tissue mechanics by laser ablation. The next two chapters provide methods for analyzing transcription followed by protocols to study growth, metabolism, ageing, and behavior in Drosophila. This volume concludes with chapters on electrophysiological recordings and methods to establish cell lines. 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 tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, Drosophila: Methods and Protocols, Second Edition is a valuable source of hands-on protocols and reviews for molecular, cell, and developmental biologists using Drosophila as model systems in their work.
Inhaltsverzeichnis
Using FlyBase: A Database of Drosophila Genes and Genomes.- The GAL4 System: A Versatile System for Manipulation and Analysis of Gene Expression.- The Q-System: A Versatile Expression System for Drosophila.- Analysis of MicroRNA Function in Drosophila.- Methods for High-Throughput RNAi screening in Drosophila Cells.- A Guide to Genome-Wide In Vivo RNAi Applications in Drosophila.- Creating Heritable Mutations in Drosophila with CRISPR-Cas9.- Performing Chromophore-Assisted Laser Inactivation in Drosophila Embryos Using GFP.- deGradFP: A System to Knockdown GFP-Tagged Proteins.- Sample Preparation and Mounting of Drosophila Embryos for Multi-View Light Sheet Microscopy.- Cultivation and Live-Imaging of Drosophila Imaginal Discs.- Cultivation and Live Imaging of Drosophila Ovaries.- Segmentation and Quantitative Analysis of Epithelial Tissues.- Laser Ablation to Probe the Epithelial Mechanics in Drosophila.- Rapid Ovary Mass-isolation (ROMi) to Obtain Large Quantities of Drosophila Egg Chambers for Fluorescent In-Situ Hybridization.- Chromatin Immunoprecipitation for Analyzing Transcription Factor Binding and Histone Modifications in Drosophila.- Protocols to Study Growth and Metabolism in Drosophila.- Protocols to Study Ageing in Drosophila.- Protocols to Study Behavior in Drosophila.- Electrophysiological Recordings from Lobula Plate Tangential Cells in Drosophila.- Methods to Establish a Drosophila Cell Line.
Zusammenfassung
This second edition volume expands on the previous edition by presenting updated protocols for several of the techniques described in the first edition of Drosophila: Methods and Protocols and current methods that cover recent breakthroughs in Drosophila research. The book begins with a description of FlyBase--a database of genes and genomes--followed by the presentation of systems for versatile gene expression in the fly. The first few chapters in this book detail gene knockdown and editing, including CRISPR-Cas9 and protein knockdown. The next few chapters are devoted to methods describing live imaging of different tissues and organs, followed by chapters on how to quantify image data and how to probe tissue mechanics by laser ablation. The next two chapters provide methods for analyzing transcription followed by protocols to study growth, metabolism, ageing, and behavior in Drosophila.This volume concludes with chapters on electrophysiological recordings and methods to establish cell lines. 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 tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, Drosophila: Methods and Protocols, Second Edition is a valuable source of hands-on protocols and reviews for molecular, cell, and developmental biologists using Drosophila as model systems in their work.
