Yeast is an ideal eukaryotic microorganism for biochemical and genetic studies, particularly since its complete genome sequence became available. The Cold Spring Harbor Laboratory Yeast Genetics Course has kept pace with the field over the last 30 years, combining essential standard methods with the latest technologies, and its course manual has followed suit. In the new 2000 edition, most of the experiments have been revised and several new experiments and techniques have been added, including the 2-hybrid system, PCR-based methods, and isolation of chromatin. This manual remains a primary source for newcomers to the field as well as the established investigator, providing complete, detailed protocols and techniques of proven robustness.

Preface Introduction Genetic Nomenclature Experiments I. Looking at Yeast Cells II. Isolation and Characterization of Auxotrophic, Temperature-sensitive, and UV-sensitive Mutants III. Meiotic Mapping IV. Mitotic Recombination and Random Spore Analysis V. Transformation of Yeast VI. Synthetic Lethal Mutants VII. Gene Replacement VIII. Isolation of ras2 Suppressors IX. Manipulating Cell Types X. Isolating Mutants by Insertional Mutagenesis XI. Two-hybrid Protein Interaction Method Techniques and Protocols 1. High-efficiency Transformation of Yeast 2. "Lazy Bones" Plasmid Transformation of Yeast Colonies 3. Yeast DNA Isolations A. Yeast DNA Miniprep (40 ml) B. Yeast DNA Miniprep (5 ml) C. A Ten-minute DNA Preparation from Yeast D. Yeast Genomic DNA: Glass Bead Preparation 4. Yeast Protein Extracts 5. Yeast RNA Isolation 6. Hydroxylamine Mutagenesis of Plasmid DNA 7. Assay of beta-Galactosidase in Yeast 8. Plate Assay for Carboxypeptidase Y 9. Random Spore Analysis 10. Yeast Vital Stains 11. Yeast Immunofluorescence 12. Actin Staining in Fixed Cells 13. PCR Protocol for PCR-mediated Gene Disruption 14. Yeast Colony PCR Protocol 15. Measuring Yeast Cell Density by Spectrophotometry 16. Cell Synchrony 17. Chromatin Immunoprecipitation 18. Flow Cytometry of Yeast DNA 19. Logarithmic Growth 20. EMS Mutagenesis 21. Tetrad Dissection 22. Making a Tetrad Dissection Needle 23. Picking Zygotes 24. Determining Plating Efficiency Appendices A. Media B. Stock Preservation C. Yeast Genetic and Physical Maps D. Templates for Making Streak Plates E. Electrophoretic Karyotypes of Strains for Southern Blot Mapping F. Strains G. Counting Yeast Cells with a Standard Hemocytometer Chamber H. Tetrad Scoring Sheet I. Trademarks J. Suppliers Related Books