Methods in yeast genetics: a cold spring harbor laboratory course manual/
David C. Amberg, Daniel J. Burke, Jeffrey N. Strathern.
- 1st ed.
- New York: Cold spring harbor lab.press, 2005.
- xvii, 230 p. : ill. ; 28 cm.
Experiments 1 Looking at Yeast Cells 2 Isolation and Charaterization of Auxotrophic, Temperature-sensitive, and Osmotic-sensitive Mutants 3 Meitotic Mapping 4 Mitotic Recombination and Random Spore Analysis 5 Transformation of Yeast 6 Synthetic Lethal Mutants 7 Gene Replacement 8 Isolation of ras2 Suppressors 9 Manipulating Cell Types 10 Isolating Mutants by Insertional Mutagenesis 11 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 Isolation 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 TAP Purification 6 Yeast RNA Isolations 7 Hydroxylamine Mutagenesis of Plansmid DNA 8 Assay of B-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 DilC5(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 Mutagensis 22 Tetrad Dissection 23 Making a Tetrad Dissection Needle 24 Picking Zygotes 25 Determining Plating Efficiency 26 DNA Miniprep from E. Coli 27 Preparing and Transfroming Competent E. Coli 28 Storing and Handling the Systematic Deletion Collection