Drosophila: Methods and Protocols

About this book

Drosophila is a comprehensive collection of methods and protocols for Drosophila, one of the oldest and most commonly used model organisms in modern biology. The text presents a diverse set of techniques that range from the basic handling of flies to more complex applications, such as the generation of mutants by homologous recombination and the analysis of developing flies by live imaging. Review chapters provide concise overviews of selected experimental systems.

This is 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. Drosophila 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.

Contents

Drosophila melanogaster and the Development of Biology in the XX century.- Getting started: An overview on raising and using Drosophila.- FlyBase: A database for the Drosophila research community.- The use of P-element transposons to generate transgenic flies.- The GAL4 system: a versatile system for the expression of genes.- P-element mutagenesis.- EMS Screens: From mutagenesis to screening and mapping.- 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.- Immunolabelling of embryos.- Imaging cellular and molecular dynamics in live embryos using fluorescent proteins.- Analysis of neuromuscular junctions: histology and in vivo imaging.- Immunolabelling of imaginal discs.- Imaging Drosophila pupal wing morphogenesis.- Microscopic analysis of the adult Drosophila retina using semi thin 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.

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