C. Elegans Methods and Applications

About this book

Nonmammalian model organisms have become a cornerstone of systems biology research. Like the Rosetta Stone, which enabled modern scholars to decode ancient Egyptian hieroglyphics, model organisms enable biologists to decipher the genetic code underlying the complex physiological processes common to all life. C. elegans provides a particularly striking example of the experimental utility of model organisms. Genetic, molecular and systems biological characterization of this organism is greatly facilitated by its short life cycle and high rate reproduction, and by the ease with which it can be cultured in the laboratory. C. elegans also has a fully sequenced and well-annotated genome, which is assembled in readily accessible public databases along with virtually all other biological data on this organism. C. elegans: Methods and Applications aims to enhance the readily available methodologies for the study of C. elegans by providing overviews and concise, step-by-step descriptions of the most state-of-the-art methods currently being utilized in the field. Some of the methods presented include forward and reverse genetic analysis, data mining and comparative genomics strategies, electron and fluorescence microscopy methods, automated imaging methods for worm behavioral analysis, functional genomics strategies, and cutting-edge methods for physiological analyses. This comprehensive collection of methods, written by experts in the field, endeavors to serve as a roadmap for researchers, illustrating the type or research that is possible with C. elegans, and illuminating its potential as a vehicle for future discovery.

Contents

An Overview of C. elegans Biology Kevin Strange Comparative Genomics in C. elegans, C. briggsae and Other Caenorhabditis Species Avril Coghlan, Jason E. Stajich and Todd W. Harris WormBase: Methods for Data Mining and Comparative Genomics Todd W. Harris and Lincoln D. Stein C. elegans Deletion Mutant Screening Robert J. Barstead and Donald G. Moerman Insertional Mutagenesis in Caenorhabditis elegans Using the Drosophila Transposon Mos1: A Method for the Rapid Identification of Mutated Genes Jean-Louis Bessereau SNP Mapping M. Wayne Davis and Marc Hammarlund Creation of Transgenic Lines Using Microparticle Bombardment Methods Vida Praitis Construction of Plasmids for RNAi and In Vitro Transcription of dsRNA Lisa Timmons Delivery Methods for RNAi in Caenorhabditis elegans Lisa Timmons Functional Genomic Approaches in C. elegans Todd Lamitina Assays for Toxicity Studies in C. elegans with Bt Crystal Proteins Larry J. Bischof, Danielle L. Huffman and Raffi V. Aroian Fluorescent Reporter Methods Harald Hutter Electrophysiological Analysis of Neuronal and Muscle Function in C. elegans Michael M. Francis and Andres Villu Maricq Sperm and Oocyte Isolation Methods for Biochemical and Proteomic Analysis Michael A. Miller Preservation of C. elegans Tissue via High-Pressure Freezing and Freeze-Substitution for Ultrastructural Analysis and Immuno-Cytochemistry Robby M. Weimer Intracellular pH Measurements In Vivo Using Green Fluorescent Protein Variants Keith Nehrke Automated Imaging of C. elegans Behavior Christopher J. Cronin, Zhaoyang Feng and William R. Schafer Intracellular Ca2+ Imaging in Caehorhabditis elegans Rex A. Kerr and William R. Schafer In Vitro Culture of C. elegans Somatic Cells< Kevin Strange and Rebecca Morrison Techniques for Analysis, Sorting, and Dispensing of C. elegans on the COPAS Flow Sorting System Rock Pulak et al.

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