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Full Description
Yeast Protocols, Third Edition presents up-to-date advances in research using yeasts as models. Chapters cover topics such as basic protocols in yeast culture and genomic manipulation, protocols that study certain organelles such as mitochondria and peroxisomes and their functions in autophagy and assays commonly used in yeast-based studies that can be adapted to other organisms. As the first sequenced living organism, budding yeast S. cerevisiae and other model yeasts have helped greatly in life science research. The easy switch between the haploid and diploid state makes yeast a paradigm of genetic manipulation. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Yeast Protocols, Third Edition seeks to serve both professionals and novices with newly-developed protocols to study this essential model organism.
Contents
Basic Investigations in Saccharomyces cerevisiae.- Isolation of Yeast Nucleic Acids.- Quantification of GFP Signals by Fluorescent Microscopy and Flow Cytometry.- Yeast Transformation by the LiAc/SS Carrier DNA/PEG Method.- Targeted Gene Deletion in Saccharomyces cerevisiae and Schizosaccharomyces pombe.- See & Catch Method for Studying Protein Complexes in Yeast Cells: A Technique Unifying Fluorescence Microscopy and Mass Spectrometry.- Yeast Two Hybrid Liquid Screening.- Targeted Mutagenesis of a Specific Gene in Yeast.- Fluorescence Staining of Mitochondria for Morphology Analysis in S. cerevisiae.- Assays for Autophagy I: the Cvt Pathway and Non-Selective Autophagy.- Assays for Autophagy II: Mitochondrial Autophagy.- Biochemically Characterizing the Subcellular Localization of Peroxisomal Proteins by Fractionation, Protease Protection and Carbonate Extraction.- Yeast Survival and Growth Assays.- Spontaneous Mutagenesis Assay.- Mutational Specificity Analysis: Assay for Mutations in the Yeast SUP4-o Gene.- piggyBac Transposon-Based Insertional Mutagenesis for the Fission Yeast Schizosaccharomyces pombe.- Replicative and Chronological Life Span Assays.- Detection of Protein Arginine Methylation in Saccharomyces cerevisiae.- Detection of Protein Post-translational Modifications from Whole Cell Extracts in Saccharomyces cerevisiae.- Ligand Engineering Using Yeast Surface Display.- Screening Combinatorial Libraries of Cyclic Peptides Using the Yeast Two-Hybrid Assay.
