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This detailed new edition provides a guide for implementing a selection of novel techniques utilizing the pioneering organism, Caenorhabditis elegans, a vital discovery engine for the rules of life, from molecules and genes, through cellular physiology, metabolism and aging, neurobiology and behavior, to individuality and interaction with the environment. Beginning with two chapters offering practical advice for setting up research programs with C. elegans, either for a new research group or for a short-term project such as a teaching laboratory, the volume continues with numerous hands-on methodologies. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, C. elegans: Methods and Applications, Third Edition offers the kind of clear guidance that will serve all researchers working with these vital and dynamic organisms.
List of contents
Starting a C. elegans Research Laboratory: Practical Advice.- The Basics of Setting Up Successful Teaching Labs and Short-Term Projects with C. elegans.- Cryopreservation of C. elegans and Other Nematodes with Dimethyl Sulfoxide and Trehalose.- Genetic Methods for Cellular Manipulation in C. elegans.- Observing and Quantifying Fluorescent Reporters.- Microbial Rhodopsin Optogenetic Tools: Application for Analyses of Synaptic Transmission and of Neuronal Network Activity in Behavior.- Optogenetic Perturbation of Individual C. elegans Pharyngeal Neurons while Monitoring Feeding Behavior.- Antibody Staining for Nematodes with Heat-Induced Antigen Retrieval (HIAR).- ExCel: Super-Resolution Imaging of C. elegans with Expansion Microscopy.- A Fusion-PCR Method for Expressing Genetic Tools in C. elegans.- Approaches for CRISPR/Cas9 Genome Editing in C. elegans.- Targeted and Random Transposon-Assisted Single-Copy Transgene Insertionin C. elegans.- Mutation Mapping and Identification by Whole-Genome Sequencing.- Lipid Extraction and Analysis.- Isolating Caenorhabditis elegans from the Natural Habitat.- Microfluidic Devices for Behavioral Analysis, Microscopy, and Neuronal Imaging in Caenorhabditis elegans.- Neuronal Microsurgery with an Yb-Doped Fiber Femtosecond Laser.- An Imaging System for Monitoring C. elegans Behavior and Aging.- Methods for Modulating and Measuring Neuromuscular Exertion in C. elegans.- Recording and Quantifying C. elegans Behavior.- Primer on Mathematical Modeling in C. elegans.
