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Methods in yeast genetics : a Cold Spring Harbor Laboratory course manual / David C. Amberg, Daniel Burke, Jeffrey Strathern.

By: Contributor(s): Publication details: Cold Spring Harbor, N.Y. : Cold Spring Harbor Laboratory Press, c2005.Edition: 2005 editionDescription: xvii, 230 p. : ill. (some col.) ; 28 cmISBN:
  • 0879697288 (pbk. : alk. paper)
Subject(s): NLM classification:
  • QW 25 A491m 2005
Online resources:
Contents:
Contents Introduction Genetic Nomenclature Experiments I. Looking at Yeast Cells II. Isolation and Characterization of Auxotrophic, Temperature- sensitive, and Osmotic-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. Isolation of Separation of Function Mutants by Two-hybrid Differential Interaction Screening Techniques and Protocols 1. High-efficiency Transformation of Yeast 2. Quick and Dirty 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 E. Yeast Genomic DNA by Spheroplasting 4. Yeast Protein Extracts 5. TAP Tag Protein Purification 6. Yeast RNA Isolation 7. Hydroxylamine Mutagenesis of Plasmid DNA 8. Assay of §-Galactosidase in Yeast 9. Plate Assay for Carboxypeptidase Y 10. Random Spore Analysis 11. Yeast Vital Stains A. DAPI Stain of Nuclear and Mitochondrial DNA B. Visualizing Mitochondria with DiOC6 or DiIC5(3) C. Visualizing Vacuoles and Endocytic Compartments with FM4-64. D. Calcofluor Staining of Chitin and Bud Scars 12. Yeast Immunofluorescence 13. Actin Staining in Fixed Cells 14. PCR-Mediated Gene Disruption A. One-Step PCR Gene Disruption. B. Gene Disruption by Double-Fusion PCR 15. Yeast Colony PCR 16. Measuring Yeast Cell Density by Spectrophotometry 17. Cell Synchrony 18. Chromatin Immunoprecipitation 19. Flow Cytometry of Yeast DNA 20. Logarithmic Growth 21. EMS Mutagenesis 22. Tetrad Dissection 23. Making a Tetrad Dissection Needle 24. Picking Zygotes 25. Determining Plating Efficiency 26. DNA Mini-Prep from E. coli 27. Preparing and Transforming Competent E. coli 28. Storing and handling of the systematic deletion collection. 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
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Book Reference Book Reference Health Sciences Library QW 25 A491m 2005 (Browse shelf(Opens below)) 1 Not for loan MBAL22010869

Includes bibliographical references and index.

Contents

Introduction

Genetic Nomenclature

Experiments
I. Looking at Yeast Cells
II. Isolation and Characterization of Auxotrophic, Temperature-
sensitive, and Osmotic-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. Isolation of Separation of Function Mutants by Two-hybrid Differential Interaction Screening

Techniques and Protocols
1. High-efficiency Transformation of Yeast
2. Quick and Dirty 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
E. Yeast Genomic DNA by Spheroplasting
4. Yeast Protein Extracts
5. TAP Tag Protein Purification
6. Yeast RNA Isolation
7. Hydroxylamine Mutagenesis of Plasmid DNA
8. Assay of §-Galactosidase in Yeast
9. Plate Assay for Carboxypeptidase Y
10. Random Spore Analysis
11. Yeast Vital Stains
A. DAPI Stain of Nuclear and Mitochondrial DNA
B. Visualizing Mitochondria with DiOC6 or DiIC5(3)
C. Visualizing Vacuoles and Endocytic Compartments with FM4-64.
D. Calcofluor Staining of Chitin and Bud Scars
12. Yeast Immunofluorescence
13. Actin Staining in Fixed Cells
14. PCR-Mediated Gene Disruption
A. One-Step PCR Gene Disruption.
B. Gene Disruption by Double-Fusion PCR
15. Yeast Colony PCR
16. Measuring Yeast Cell Density by Spectrophotometry
17. Cell Synchrony
18. Chromatin Immunoprecipitation
19. Flow Cytometry of Yeast DNA
20. Logarithmic Growth
21. EMS Mutagenesis
22. Tetrad Dissection
23. Making a Tetrad Dissection Needle
24. Picking Zygotes
25. Determining Plating Efficiency
26. DNA Mini-Prep from E. coli
27. Preparing and Transforming Competent E. coli
28. Storing and handling of the systematic deletion collection.
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

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