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.
Gespeichert in:
| Hauptverfasser: | , , |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
Cold Spring Harbor, NY
Cold Spring Harbor Laboratory Press
2005
|
| Ausgabe: | 2005 ed. |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Zusammenfassung: | "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. |
| Beschreibung: | Früher u.d.T.: Methods in yeast genetics |
| Beschreibung: | XVII, 230 S. Ill. |
| ISBN: | 0879697288 |
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Datensatz im Suchindex
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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
|
| adam_txt |
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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| spelling | Amberg, David C. Verfasser aut Methods in yeast genetics a Cold Spring Harbor Laboratory Course Manual David C. Amberg ; Daniel J. Burke ; Jeffrey N. Strathern 2005 ed. Cold Spring Harbor, NY Cold Spring Harbor Laboratory Press 2005 XVII, 230 S. Ill. txt rdacontent n rdamedia nc rdacarrier Früher u.d.T.: Methods in yeast genetics "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. 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 rswk-swf Genetik (DE-588)4071711-2 gnd rswk-swf Labortechnik (DE-588)4123602-6 gnd rswk-swf Saccharomyces cerevisiae (DE-588)4178812-6 s Genetik (DE-588)4071711-2 s Labortechnik (DE-588)4123602-6 s b DE-604 Burke, Daniel J. Verfasser aut Strathern, Jeffrey N. 19XX- Verfasser (DE-588)1236310853 aut HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=014277670&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| 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_exact_search_txtP | 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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