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Yeast Protocols contains many key techniques for studying the biology of yeasts at both the cellular and molecular levels. Working primarily from Saccharomyces cerevisiae, the expert contributors explain step-by-step how to successfully isolate, identify, and culture yeasts; the secrets of meiotic mapping; how to use PFGE in karyotyping and gene localization; the methods for purification and analysis of various cell components; and, the construction and exploitation of genomic DNA clone banks. They also cover the latest methods for chromosome engineering, insertional mutagenesis by Ty elements, mRNA abundance and half-life measurements, the use of reporter gene systems, genotoxicity testing, and more. "Yeast Protocols" follows the widely applauded Humana Methods in Molecular Biology style: brief introductions putting the particular method in context, comprehensive lists of materials, cookbook style instructions, and troubleshooting notes to avoid common pitfalls and solve problems. The techniques can be used with confidence and success by both inexperienced newcomers and established researchers.
Isolation and identification of Yeasts from Natural Habitats. Maintenance and Culture of Yeasts. Mutagenesis in Yeast. Rare-Mating and Cytoduction in Saccharomyces cerevisiae. Protoplast Fusion in Saccharomyces cerevisiae. Meiotic Analysis. Automated Ascus Dissection. Karyotyping of Yeast Strains by PFGE. Chromosomal Localization of Genes by PFGE Techniques. Isolation, Purification, and Analysis of Nuclear DNA in Yeast Taxonomy. Isolation of Yeast DNA. Isolation of Mitochondrial DNA. Isolation of Yeast Plasma Membranes. The Extraction and Analysis of Sterols from Yeast. Peroxisome Isolation. Transformation of Lithium-Treated Yeast Cells and the Selection of Auxotrophic and Dominant Markers. Biolistic Transformation of Yeasts. Genomic Yeast DNA Clone Banks. Yeast Colony Hybridization. Plasmid Copy Number by A Simple Hybridization Method. Determination of Plasmid Copy Number in Yeast. Determination of Chromosome Ploidy in Yeast. Chromosome Engineering in Yeast with a Site-specific Recombination System from a Heterologous Yeast Plasmid. Insertional Mutagenesis by Ty Elements in Saccharomyces cerevisiae. Reporter Gene Systems for Assaying Gene Expression in Yeast. Preparation and Use of Yeast Cell-Free Translation Lysate. Electrophoretic Analysis of Yeast Proteins. Preparation of Total RNA. mRNA Abundance and Half-Life Measurements. Polysome Analysis. Induction of Heat Shock Proteins and Thermotolerance. Rhodamine B Assay for Estimating Activity of Killer Toxins Permeabilizing Cytoplasmic Membranes. Nystalin-Rhodamine B Assay for Estimating Activity of Killer Toxin from Kluyveromyces lactis. Application of Killer Toxins in Stepwise Selection of Hybrids and Cybrids Obtained by Induced Protoplast Fusion. Killer Plaque Technique for Selecting Hybrids and Cybrids Obtained by Induced Protoplast Fusion. Genotoxicity Testing in Schizosaccharomyces pombe. Purification and Quantification of Saccharomyces cerevisiae Cytochrome P450. Calorimetry of Whole Yeast Cells. Light Microscopy Methods. Fluorescence Microscopy Methods. Immunoelectronmicroscopy of Yeast. Index.
