Read more
Drosophila melanogaster has been the model system of choice for many inv- tigators over the past hundred years. Due to its long-standing tradition as a model organism, many techniques used in Drosophila have been established and c- tinue to be developed to answer a variety of scientific questions. The recent inv- tion of techniques allowing the knock-down of genes by RNA interference and gene replacement by homologous recombination are two important demonst- tions of the ongoing efforts to broaden the technical repertoire at hand for researchers working with Drosophila. This volume is a collection of protocols covering both standard techniques as well as novel methods. An introductory chapter highlights the importance of Drosophila as a model system for the dev- opment of Biology in the 20th century and review chapters provide concise and up-to-date overviews on selected experimental systems. This book makes no attempt to be comprehensive. A number of frequently used standard techniques were selected and are described in-depth to allow novices to get started with Drosophila. In addition, detailed protocols of recently developed methods that we felt will become of broad use in the Drosophila community within the next years are included. I am grateful to Konrad Basler, Suzanne Eaton, Elisabeth Knust, Pavel Tomancak, and Andreas Wodarz for alerting me to novel techniques and for their advice on selecting the protocols for this book. I would also like to thank all the authors for their expert contribution to this Volume. Christian Dahmann v Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .
List of contents
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.
Summary
Drosophila melanogaster has been the model system of choice for many inv- tigators over the past hundred years. Due to its long-standing tradition as a model organism, many techniques used in Drosophila have been established and c- tinue to be developed to answer a variety of scientific questions. The recent inv- tion of techniques allowing the knock-down of genes by RNA interference and gene replacement by homologous recombination are two important demonst- tions of the ongoing efforts to broaden the technical repertoire at hand for researchers working with Drosophila. This volume is a collection of protocols covering both standard techniques as well as novel methods. An introductory chapter highlights the importance of Drosophila as a model system for the dev- opment of Biology in the 20th century and review chapters provide concise and up-to-date overviews on selected experimental systems. This book makes no attempt to be comprehensive. A number of frequently used standard techniques were selected and are described in-depth to allow novices to get started with Drosophila. In addition, detailed protocols of recently developed methods that we felt will become of broad use in the Drosophila community within the next years are included. I am grateful to Konrad Basler, Suzanne Eaton, Elisabeth Knust, Pavel Tomancak, and Andreas Wodarz for alerting me to novel techniques and for their advice on selecting the protocols for this book. I would also like to thank all the authors for their expert contribution to this Volume. Christian Dahmann v Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .
