Drosophila Protocols

This exceptional laboratory manual describes thirty-seven procedures most likely to be used in the next decade for molecular, biochemical, and cellular studies on Drosophila. They were selected after extensive consultation with the research community and rigorously edited for clarity, uniformity, and conciseness.The methods included permit investigation of chromosomes, cell biology, molecular biology, genomes, biochemistry, and development. Each protocol includes the basic information needed by novices, with sufficient detail to be valuable to experienced investigators. Each method is carefully introduced and illustrated with figures, tables, illustrations, and examples of the data obtainable. The book's appendices include key aspects of Drosophila biology, essential solutions, buffers, and recipes.An evolution of Michael Ashburner's 1989 classic Drosophila: A Laboratory Manual, this book is an essential addition to the personal library of Drosophila investigators and an incomparable resource for other research groups with goals likely to require fly-based technical approaches.

Leading drosophilists describe in step-by-step detail all the essential techniques for studying Drosophila chromosomes and suggest new avenues for scientific exploration. The chapters emphasize specimen preparation (from dissection to mounting) and cover both polytene and mitotic/meiotic chromosomes in depth. Each fully tested and readily reproducible protocol offers a background introduction, equipment and reagent lists, and tips on troubleshooting and avoiding pitfalls. A cutting-edge FISH and immunolocalization technique will be important for discovering how DNA sequence influences higher-order chromosome architecture and ultimately gene expression.

This third edition expands on the previous editions by providing updated protocols and current methods that cover recent breakthroughs in Drosophila research. Chapters guide readers through FlyBase, versatile gene expression systems, analysis of microRNA function, single-cell transcriptome data and metabolism, recent applications of CRISPR for precise genome editing, transcriptional activation and cell lineage tracing, protein inhibition and tagging, optogenetic and optochemical control of tissue mechanics, AFM measurements, sample preparation for electron microscopy, live imaging of different tissues and organs, and quantitative image analysis. Written in the format of the highly successful Methods in Molecular Biology series, chapters include an introduction to their topic, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls Authoritative and cutting-edge, Drosophila: Methods and Protocols, Third Edition serves as a useful and practical guide to new researchers and experts using Drosophila as a model system. Chapter 7 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

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.

This detailed volume compiles numerous methods to explore fly oogenesis. Beginning with updated protocols from isolating and staining the ovary to numerous imaging techniques and genetic protocols for cell-specific assessment and CRISPR-mediated mutagenesis, the volume continues by detailing techniques from assessing cytoskeletal structures to uncovering protein-protein interactions, with closing chapters exploring how Drosophila oogenesis can be used in the classroom and in outreach programs to increase interest in biomedical research, STEM education, and STEM careers. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Drosophila Oogenesis: Methods and Protocols serves as an ideal guide for both new and expert researchers working with this robust and versatile model organism.

Based on Cold Spring Harbor Laboratory's long-running course, Drosophila Neurobiology: A Laboratory Manual offers detailed protocols and background material for researchers interested in using Drosophila as an experimental model for investigating the nervous system. This manual covers three approaches to the field: analysis of neural development, recording and imaging activities in the nervous system, and analysis of behavior. Techniques described include molecular, genetic, electrophysiological, imaging, behavioral and developmental methods.