Methods in Yeast Genetics 2000: A Cold Spring Harbor Laboratory Course Manual

by Burke, Dan; Dawson, Dean; Stearns, Tim; Cold Spring Harbor Laboratory

• ISBN13: 9780879695880
• ISBN10: 0879695889
• Additional ISBN(s): 9780879697280
• Edition: Lab
• Format: Paperback
• Copyright: 2000-08-01
• Publisher: Cold Spring Harbor Laboratory Pr
• More Book Details
• Purchase Benefits

Summary

Revised edition of Methods in Yeast Genetics, 1997 Edition, c1998. Focuses on the baker's yeast, Saccharomyces cerevisiae. Readers should be able to carry out all of the techniques commonly employed by yeast geneticists and follow the literature with ease. Softcover.

Table of Contents

Preface	vii
Introduction	ix
Genetic Nomenclature	xiii
Experiments
Looking at Yeast Cells	1	(6)
Isolation and Characterization of Auxotrophic, Temperaturesensitive, and UV-sensitive Mutants	7	(10)
Meiotic Mapping	17	(12)
Mitotic Recombination and Random Spore Analysis	29	(10)
Transformation of Yeast	39	(10)
Synthetic Lethal Mutants	49	(6)
Gene Replacement	55	(18)
Isolation of ras2 Suppressors	73	(6)
Manipulating Cell Types	79	(10)
Isolating Mutants by Insertional Mutagenesis	89	(8)
Two-hybrid Protein Interaction Method	97	(6)
Techniques and Protocols
High-efficiency Transformation of Yeast	103	(4)
``Lazy Bones'' Plasmid Transformation of Yeast Colonies	107	(2)
Yeast DNA Isolations	109	(6)
Yeast DNA Miniprep (40 ml)	109	(1)
Yeast DNA Miniprep (5 ml)	110	(1)
A Ten-minute DNA Preparation from Yeast	111	(2)
Yeast Genomic DNA: Glass Bead Preparation	113	(2)
Yeast Protein Extracts	115	(2)
Yeast RNA Isolation	117	(4)
Hydroxylamine Mutagenesis of Plasmid DNA	121	(2)
Assay of β-Galactosidase in Yeast	123	(4)
Plate Assay for Carboxypeptidase Y	127	(2)
Random Spore Analysis	129	(2)
Yeast Vital Stains	131	(2)
Yeast Immunofluorescence	133	(4)
Actin Staining in Fixed Cells	137	(2)
PCR Protocol for PCR-mediated Gene Disruption	139	(2)
Yeast Colony PCR Protocol	141	(2)
Measuring Yeast Cell Density by Spectrophotometry	143	(2)
Cell Synchrony	145	(4)
Chromatin Immunoprecipitation	149	(6)
Flow Cytometry of Yeast DNA	155	(2)
Logarithmic Growth	157	(2)
EMS Mutagenesis	159	(2)
Tetrad Dissection	161	(4)
Making a Tetrad Dissection Needle	165	(2)
Picking Zygotes	167	(2)
Determining Plating Efficiency	169	(36)
Appendices
A. Media	171	(12)
B. Stock Preservation	183	(2)
C. Yeast Genetic and Physical Maps	185	(8)
D. Templates for making Streak Plates	193	(2)
E. Electrophoretic Karyotypes of Strains for Southern Blot Mapping	195	(2)
F. Strains	197	(2)
G. Counting Yeast Cells with a Standard Hemocytometer Chamber	199	(2)
H. Tetrad Scoring Sheet	201	(2)
I. Trademarks	203	(2)
J. Suppliers	205