This is a comprehensive collection of methods and protocols for Drosophila, one of the oldest and most commonly used model organisms in modern biology. The protocols are written by the scientists who invented the methods making this book the perfect reference manual for Drosophila researchers. The straightforward presentation of Drosophila protocols will bolster successful experimental design and lead to reproducible results, and the troubleshooting tips are an invaluable aid for researchers at all levels. This book will advance the use of Drosophila as a model organism by collecting standard, introductory protocols and advanced protocols, simultaneously providing a platform for introductory research and facilitation advanced study. The text presents a diverse set of techniques that range from the basic handling of flies to more complex applications. Review chapters provide concise overviews of selected experimental systems.

Protocols are written by the scientists who invented the methods Includes methods for homologous recombination in Drosophila Includes methods for live imaging in embryos, larvae, and pupae

Klappentext

With its long-standing tradition as a model organism, many techniques using Drosophila melanogaster have been established and continue to be developed, while the recent invention of techniques allowing the knock-down of genes by RNA interference and gene replacement by homologous recombination demonstrates the ongoing efforts to broaden the technical repertoire. In Drosophila: Methods and Protocols, leading experts contribute a collection of protocols covering standard techniques as well as novel methods. As a volume in the highly successful Methods in Molecular Biology™ series, the chapters contain brief introductions to the subject matter, 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 essential, Drosophila: Methods and Protocols allows novices and experts alike to tap into the power of one hundred years of expertise involving this vital model system.

Inhalt
Drosophila melanogaster and the Development of Biology in the 20th Century.- Getting Started.- FlyBase.- The Use of P-Element Transposons to Generate Transgenic Flies.- The GAL4 System.- P-Element Mutagenesis.- EMS Screens.- RNAi Screening in Cultured Drosophila Cells.- Methods for Homologous Recombination in Drosophila.- Recombinases and Their Use in Gene Activation, Gene Inactivation, and Transgenesis.- Cuticle Preparation of Drosophila Embryos and Larvae.- Immunolabeling of Embryos.- Imaging Cellular and Molecular Dynamics in Live Embryos Using Fluorescent Proteins.- Analysis of Neuromuscular Junctions.- Immunolabeling of Imaginal Discs.- Imaging Drosophila Pupal Wing Morphogenesis.- Microscopic Analysis of the Adult Drosophila Retina Using Semithin Plastic Sections.- Fluorescent In Situ Hybridization Protocols in Drosophila Embryos and Tissues.- Linear RNA Amplification for the Production of Microarray Hybridization Probes.- An Overview of the Identification, Detection, and Functional Analysis of Drosophila MicroRNAs.- Extraction and Immunoblotting of Proteins From Embryos.- Purification of Drosophila Protein Complexes for Mass Spectrometry.- Mass Production of Drosophila Embryos and Chromatographic Purification of Native Protein Complexes.- Flow Cytometric Analysis of Drosophila Cells.- Drosophila Cell Lines as Model Systems and as an Experimental Tool.