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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.
List of contents
The Chromosomes of Drosophila melanogaster.- Spermatogenesis.- Time-Lapse Imaging of Male Meiosis by Phase-Contrast and Fluorescence Microscopy.- Immunocytological Analysis of Oogenesis.- Cytological Analysis of Oogenesis.- Polytene Chromosomes From Ovarian Nurse Cells of Drosophila melanogaster otu Mutants.- Feulgen-DNA Cytophotometry for Estimating C Values.- Fluorescent BrdU Labeling and Nuclear Flow Sorting of the Drosophila Ovary.- Studying Nuclear Organization in Embryos Using Antibody Tools.- Whole-Mount Fluorescence In Situ Hybridization to Chromosomes of Embryos.- Orcein Staining and the Identification of Polytene Chromosomes.- Salivary Chromosome Analysis of Aberrations.- In Situ Hybridization to Polytene Chromosomes.- Combined Immunostaining and FISH Analysis of Polytene Chromosomes.- Electron Microscopy of Polytene Chromosomes.- Analysis of Mitosis in Squash Preparations of Larval Brains.- Analyzing Chromosome Function by High Frequency Formation of Dicentric Chromosomes In Vivo.- Analysis of Chromosomes of the Larval CNS by FISH and BrdU Labeling.- Immunostaining of Squash Preparations of Chromosomes of Larval Brains.- Visualizing Mitosis in Whole-Mount Larval Brains.- Immunostaining of Whole-Mount Imaginal Discs.- Wing Somatic Mutation and Recombination Test.- Analysis of Histoblasts.- Visualizing Apoptosis.- RNAi in Cultured Drosophila Cells.
Summary
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.
Additional text
...this volume will be worth returning to for years to come...the spirit of this volume is not that it will get you through, but that it will make you comfortable with the technology and put you in touch with the art form...a valuable and stimulating source of essential technical information for all experimental 'drosophilists'. - Nature Genetics
