Methods in yeast genetics a Cold Spring Harbor Laboratory Course Manual

"Methods in Yeast Genetics" is a course that has been offered annually at Cold Spring Harbor for the last 30 years. This provides a set of teaching experiments along with the protocols and recipes for the standard techniques and reagents used in the study of yeast biology.

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Hauptverfasser: Amberg, David C. (VerfasserIn), Burke, Daniel J. (VerfasserIn), Strathern, Jeffrey N. 19XX- (VerfasserIn)
Format: Buch
Sprache:English
Veröffentlicht: Cold Spring Harbor, NY Cold Spring Harbor Laboratory Press 2005
Ausgabe:2005 ed.
Schlagworte:
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adam_text Titel: Methods in yeast genetics Autor: Amberg, David C Jahr: 2005 Contents Preface vii Introduction ix Genetic Nomenclature xiii Experiments I. Looking at Yeast Cells 1 II. Isolation and Characterization of Auxotrophic, Temperature-sensitive, and Osmotic-sensitive Mutants 11 III. Meiotic Mapping 21 IV. Mitotic Recombination and Random Spore Analysis 33 V. Transformation of Yeast 43 VI. Synthetic Lethal Mutants 53 VII. Gene Replacement 59 VIII. Isolation of ras2 Suppressors 77 IX. Manipulating Cell Types 85 X. Isolating Mutants by Insertional Mutagenesis 95 XI. Isolation of Separation of Function Mutants by Two-hybrid Differential Interaction Screening 101 Techniques and Protocols 1. High-efficiency Transformation of Yeast 109 2. Quick and Dirty Plasmid Transformation of Yeast Colonies 113 3. Yeast DNA Isolations 115 A. Yeast DNA Miniprep (40 ml) 115 B. Yeast DNA Miniprep (5 ml) 116 C. A Ten-minute DNA Preparation from Yeast 117 D. Yeast Genomic DNA: Glass-bead Preparation 119 4. Yeast Protein Extracts 121 5. TAP Purification 123 6. Yeast RNA Isolations 127 7. Hydroxylamine Mutagenesis of Plasmid DNA 133 8. Assay of p-Galactosidase in Yeast 135 V Vi a CONTENTS 9. Plate Assay for Carboxypeptidase Y 141 10. Random Spore Analysis . 143 11. Yeast Vital Stains 145 A. DAPI Stain of Nuclear and Mitochondrial DNA 145 B. Visualizing Mitochondria with DiOC6 or DiICs(3) 145 C. Visualizing Vacuoles and Endocytic Compartments with FM4-64 146 D. Calcofluor Staining of Chitin and Bud Scars 146 12. Yeast Immunofluorescence 149 13. Actin Staining in Fixed Cells 153 14. PCR-mediated Gene Disruption 155 A. One-step PCR Gene Disruption 155 B. Gene Disruption by Double-fusion PCR 157 15. Yeast Colony PCR 161 16. Measuring Yeast Cell Density by Spectrophotometry 163 17. Cell Synchrony 165 18. Chromatin Immunoprecipitation 169 19. Flow Cytometry of Yeast DNA 175 20. Logarithmic Growth 177 21. EMS Mutagenesis 179 22. Tetrad Dissection 181 23. Making a Tetrad Dissection Needle 185 24. Picking Zygotes 187 25. Determining Plating Efficiency 189 26. DNA Miniprep from E. coli 191 27. Preparing and Transforming Competent E. coli 193 28. Storing and Handling the Systematic Deletion Collection 197 199 Appendices A. Media B. Stock Preservation 211 C. Yeast Genetic and Physical Maps 213 D. Templates for Making Streak Plates 221 E. Electrophoretic Karyotypes of Strains for Southern Blot Mapping 223 F. Strains 225 G. Counting Yeast Cells with a Standard Hemocytometer Chamber 227 H. Tetrad Scoring Sheet 229
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spellingShingle Amberg, David C.
Burke, Daniel J.
Strathern, Jeffrey N. 19XX-
Methods in yeast genetics a Cold Spring Harbor Laboratory Course Manual
Fungos (genética) larpcal
Genetica gtt
Gist gtt
Levure - Génétique - Manuels de laboratoire
Yeast fungi Genetics Laboratory manuals
Yeasts genetics Laboratory Manuals
Saccharomyces cerevisiae (DE-588)4178812-6 gnd
Genetik (DE-588)4071711-2 gnd
Labortechnik (DE-588)4123602-6 gnd
subject_GND (DE-588)4178812-6
(DE-588)4071711-2
(DE-588)4123602-6
title Methods in yeast genetics a Cold Spring Harbor Laboratory Course Manual
title_auth Methods in yeast genetics a Cold Spring Harbor Laboratory Course Manual
title_exact_search Methods in yeast genetics a Cold Spring Harbor Laboratory Course Manual
title_full Methods in yeast genetics a Cold Spring Harbor Laboratory Course Manual David C. Amberg ; Daniel J. Burke ; Jeffrey N. Strathern
title_fullStr Methods in yeast genetics a Cold Spring Harbor Laboratory Course Manual David C. Amberg ; Daniel J. Burke ; Jeffrey N. Strathern
title_full_unstemmed Methods in yeast genetics a Cold Spring Harbor Laboratory Course Manual David C. Amberg ; Daniel J. Burke ; Jeffrey N. Strathern
title_short Methods in yeast genetics
title_sort methods in yeast genetics a cold spring harbor laboratory course manual
title_sub a Cold Spring Harbor Laboratory Course Manual
topic Fungos (genética) larpcal
Genetica gtt
Gist gtt
Levure - Génétique - Manuels de laboratoire
Yeast fungi Genetics Laboratory manuals
Yeasts genetics Laboratory Manuals
Saccharomyces cerevisiae (DE-588)4178812-6 gnd
Genetik (DE-588)4071711-2 gnd
Labortechnik (DE-588)4123602-6 gnd
topic_facet Fungos (genética)
Genetica
Gist
Levure - Génétique - Manuels de laboratoire
Yeast fungi Genetics Laboratory manuals
Yeasts genetics Laboratory Manuals
Saccharomyces cerevisiae
Genetik
Labortechnik
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