Methods in yeast genetics : a Cold Spring Harbor Laboratory course manual

Experiments: looking at yeast cells isolation and characterization of auxotrophic, temperature-sensitive and UV-sensitive mutants meiotic mapping mitotic recombination and random spore analysis transformation of yeast synthetic lethal mutants gene replacement isolation of ras2 suppressors manipulating cell types isolating mutants by insertional mutagenesis two-hybrid protein interaction method. Techniques and protocols: high-efficiency transformation of yeast Lazy Bones plasmid transformation of yeast colonies yeast DNA isolations yeast DNA miniprep (40 ml) yeast DNA miniprep (5 ml) a ten-minute DNA preparation from yeast yeast genomic DNA - glass bead preparation yeast protein extracts yeast RNA isolation hydroxylamine mutagenesis of plasmid DNA assay of beta-galactosidase in yeast plate assay for carboxypeptidase Y random spore analysis yeast vital stains yeast immunofluorescence actin staining in fixed cells PCR protocol for PCR-mediated gene disruption yeast colony PCR protocol measuring yeast cell density by spectrophotometry cell synchrony chromatin immunoprecipitation flow cytometry of yeast DNA logarithmic growth EMS mutagenesis tetrad dissection making a tetrad dissection needle picking zygotes determining plating efficiency. Appendices: media stock preservation yeast genetic and physical maps templates for making streak plates electrophoretic karyotypes of strains for southern blot mapping strains counting yeast cells with a standard hemocytometer chamber tetrad scoring sheet trademarks suppliers.