Table of Contents
Preface
Acknowledgments
Yeast Strains
Yeast Plasmids
Yeast Nomenclature
Plates and Media
A Few Words on Sterile Technique
Yeast Resources
EXPERIMENTS
I. Transformation of Plasmids and Integrating DNAs
II. Looking at Yeast Cells
III. Manipulating Mating-Type and Epigenetic Transcriptional Silencing
IV. Mating, Meiosis, and Tetrad Dissection
V. Isolation and Characterization of Auxotrophic and Temperature-Sensitive Mutants
VI. Working with Essential Genes
VII. Synthetic Lethal Mutants and Random Sporulation
VIII. Measuring Mutation Rates and Studying Human Genetic Variation in Yeast
IX. Mutation Detection Using Comparative Genomic Hybridization
X. Mutation Detection Using Whole-Genome Sequencing and Linkage
XI. Barcode Sequencing and Comparative Functional Genomics
TECHNIQUES AND PROTOCOLS
1. Genomic Modifications with PCR Products
2. High-Efficiency Yeast Transformation
3. Using the C6 Cytometer
4. Modified Hoffman–Winston Genomic DNA Preparation
5. Indirect Immunofluorescence Microscopy
6. Yeast Vital Stains
7. Actin Staining in Fixed Yeast Cells
8. Preparation of Slides with Agarose Pads for Imaging of Live Yet Immobile Yeast
9. Sporulation and Tetrad Dissection
10. Making a Tetrad Dissection Needle
11. EMS Mutagenesis
12. Counting Yeast Cells with a Hemocytometer
13. Flow Cytometry of Yeast DNA Content Using SYBR Green
14. Storage and Handling of the Systematic Deletion Collection
15. Scoring SGA Screens with SGATools
16. Measuring DNA Concentration with the Qubit Fluorometer
17. Colony PCR
18. Training for the Plate Race
APPENDICES
Appendix A: Media
Rich Media
Synthetic Media
Sporulation Medium
Indicator Media
Drug Selection Media
SGA Media
Appendix B: Tetrad Dissection Sheets
Appendix C: 96-Well Plate Template
Appendix D: Templates for Making Streaks and Patches
Appendix E: General Safety and Hazardous Material Information
Index
