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Methods in Yeast Genetics and Genomics, 2015 Edition: A CSHL Course Manual


Book Series:  Cold Spring Harbor Laboratory Course Manual
Subject Area(s):  Yeast; Genetics; Laboratory Techniques

By Maitreya Dunham, University of Washington; Marc Gartenberg, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey; Grant W. Brown, University of Toronto

Download a Free Excerpt from Methods in Yeast Genetics and Genomics, 2015 Edition: A CSHL Course Manual:

Preface
Index

© 2015 • 256 pages, illustrated (21 color, 23 B&W), index
Hardcover • $89 71.20
ISBN  978-1-621821-34-2
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  •     Description    
  •     Contents    

Description

Methods in Yeast Genetics is a course that has been offered annually at Cold Spring Harbor Laboratory for the last 45 years. This is an updated edition of the course manual, which provides a set of teaching experiments, along with protocols and recipes for the standard techniques and reagents used in the study of yeast biology. Since the last edition of the manual was published (2005), revolutionary advances in genomics, proteomics, and imaging technologies have had a significant impact on the field. The 11 experiments included in this manual provide a foundation of methods for any modern-day yeast lab. These methods emphasize combinations of classical and modern genetic approaches, including isolation and characterization of mutants, two-hybrid analysis, tetrad analysis, complementation, and recombination. Also covered are molecular genetic techniques for genome engineering. Additional experiments introduce fundamental techniques in yeast genomics, including both performance and interpretation of Synthetic Genetic Array analysis, multiplexed whole genome and barcode sequencing, and comparative genomic hybridization to DNA arrays. Comparative genomics is introduced using different yeast strains to study natural variation, evolution, and quantitative traits. This manual covers the full repertoire of genetic approaches needed to dissect complex biological problems in the yeast Saccharomyces cerevisiae.

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