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Methods in Yeast Genetics

  •  ISBN-10 0-87969-588-9

  •  ISBN-13 978-0-87969-588-0

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Related Categories Books and Software, Cold Spring Harbor Laboratory Press/ Scion, Genetics Books, Labware, Manufacturer Browser More...
publication info   D. Burke, et al., Cold Spring Harbor Laboratory Press, 2000, 205 pp., soft cover
mfr. no.   Cold Spring Harbor

Description

General description

Yeast is an ideal eukaryotic microorganism for biochemical and genetic studies, particularly since its complete genome sequence became available. This Cold Spring Harbor Laboratory Yeast Genetics Course Manual combines essential standard methods with the latest technologies. In this 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.

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 Hemacytometer Chamber
H. Tetrad Scoring Sheet
I. Trademarks
J. Suppliers

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