© 2000 • 728 pp., illus., appendices, index
Paperback • $81.00 • ISBN  978-087969827-0

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 outstanding features of this protocol collection are:

• Scope: The methods included permit investigation of chromosomes, cell biology, molecular biology, genomes, biochemistry, and development.
• Depth: Each protocol includes the basic information needed by novices, with sufficient detail to be valuable to experienced investigators.
• Format: Each method is carefully introduced and illustrated with figures, tables, illustrations, and examples of the data obtainable.
• Added value: 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.

  “… whether you are a fly novice keen to test the waters or a die-hard geneticist looking to expand your horizons, if you are planning to do experiments using Drosophila over the next few years, you will find this book greatly useful.”
      —Nature Cell Biology

  “The fly community is evolving, its tastes are becoming increasingly refined and its work becomes more interdisciplinary by the year. Drosophila Protocols reflects this changing mood and I expect it will become the fly pushers' techniques bible for the coming decade.”
      —Trends in Genetics

Preface
Abbreviations

CHROMOSOMES

Chapter 1: Preparation and Analysis of Drosophila Mitotic Chromosomes (S. Pimpinelli, S. Bonaccorsi, L. Fanti, and M. Gatti)

Chapter 2: In Situ Hybridization to Somatic Chromosomes (A.F. Dernburg)

Chapter 3: BrdU Labeling of Chromosomes (A.W. Shermoen)

Chapter 4: Analysis of Meiosis of Fixed and Live Oocytes by Light Microscopy (H.J.G. Matthies, M. Clarkson, R.B. Saint, R. Namba, and R.S. Hawley)

Chapter 5: Cytological Analysis of Spermatocyte Growth and Male Meiosis in Drosophila melanogaster (S. Bonaccorsi, M.G. Giansanti, G. Cenci, and M. Gatti)

Chapter 6: Preparation and Analysis of Polytene Chromosomes (J.A. Kennison)

Chapter 7: In Situ Hybridization to Polytene Chromosomes (M.-L. Pardue)

Chapter 8: Mapping Protein Distributions on Polytene Chromosomes by Immunostaining (R. Paro)

CELL BIOLOGY

Chapter 9: Fluorescent Analysis of Drosophila Embryos (W.F. Rothwell and W. Sullivan)

Chapter 10: Imaginal Discs (S.S. Blair)

Chapter 11: The Neuromuscular Junction (H.J. Bellen and V. Budnik)

Chapter 12: Histological Techniques for the Drosophila Eye. Part I: Larva and Pupa (T. Wolff)

Chapter 13: Histological Techniques for the Drosophila Eye. Part II: Adult (T. Wolff)

Chapter 14: Preparation of Thin Sections of Drosophila for Examination by Transmission Electron Microscopy (K.L. McDonald, D.J. Sharp, and W. Rickoll)

Chapter 15: Electrophysiological Approaches to the Neuromusculature (K.S. Broadie)

Chapter 16: Functional Assays of the Peripheral and Central Nervous System (K.S. Broadie)

Chapter 17: GFP and Other Reporters (T. Hazelrigg)

Chapter 18: Quantitative Microinjection of Drosophila Embryos (D.P. Kiehart, J.M. Crawford, and R.A. Montague)

Chapter 19: Targeted Disruption of Gene Function in Drosophila by RNA Interference (L. Misquitta and B.M. Paterson)

Chapter 20: Drosophila Cell Culture and Transformation (L. Cherbas and P. Cherbas)

Chapter 21: Generating Antibodies against Drosophila Proteins (I. Rebay and R.G. Fehon)

Chapter 22: Photoactivated Gene Expression for Cell-fate Mapping and Cell Manipulation (J. Minden, R. Namba, and S. Cambridge)

MOLECULAR BIOLOGY

Chapter 23: Recovery of DNA Sequences Flanking P-element Insertions: Inverse PCR and Plasmid Rescue (A.M. Huang, E.J. Rehm, and G.M. Rubin)

Chapter 24: GAL4-mediated Ectopic Gene Expression in Drosophila (P. van Roessel and A.H. Brand)

Chapter 25: Functional Cell Ablation (S.T. Sweeney, A. Hidalgo, J.S. deBelle, and H. Keshishian)

GENOMICS

Chapter 26: Preparation of DNA from E. coli Cells Containing Bacteriophage P1 Clones (E.R. Lozovskaya)

Chapter 27: Drosophila Microarrays: From Arrayer Construction to Hybridization (K.P. White and K.C. Burtis)

Chapter 28: Using Drosophila Genome Databases (S. Misra, M.A. Crosby, and R.A. Drysdale)

Chapter 29: Preparation and Analysis of High-molecular-weight DNA in Drosophila (G.H. Karpen)

BIOCHEMISTRY

Chapter 30: Culturing Large Populations of Drosophila for Protein Biochemistry (J.C. Sisson)

Chapter 31: Preparation of Nuclear Extracts from Drosophila Embryos and In Vitro Transcription Analysis (M.J. Pazin)

Chapter 32: Preparation of Membrane Proteins from Drosophila Embryos (C.X. Zhang & T.-S. Hsieh)

Chapter 33: Preparing Cytoplasmic Extracts from Drosophila Embryos (M. Moritz)

Chapter 34: Immunoblotting of Proteins from Single Drosophila Embryos (T.T. Su)

THE ORGANISM

Chapter 35: Laboratory Culture of Drosophila (M. Ashburner and J. Roote)

Chapter 36: Preparation of Larval and Adult Cuticles for Light Microscopy (D.L. Stern and E. Sucena)

Chapter 37: Exposing Drosophila to Neuroactive Drugs (J. Hirsh)

APPENDICES

1. 1989 Table of Contents from Drosophila: A Laboratory Manual
2. Anatomical Drawings of Drosophila
3. Solutions and Reagents
4. Cautions
5. Suppliers
6. Trademarks

Index