Methods in Yeast Genetics: A Cold Spring Harbor Laboratory Course Manual / Edition 2000

Methods in Yeast Genetics: A Cold Spring Harbor Laboratory Course Manual / Edition 2000

by Tim Stearns
     
 

ISBN-10: 0879695889

ISBN-13: 9780879695880

Pub. Date: 01/01/2000

Publisher: Cold Spring Harbor Laboratory Press

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

Overview

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.

Product Details

ISBN-13:
9780879695880
Publisher:
Cold Spring Harbor Laboratory Press
Publication date:
01/01/2000
Edition description:
LAB MANUAL
Pages:
205
Product dimensions:
8.50(w) x 10.90(h) x 0.70(d)

Table of Contents

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

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