Bgal plate overlay assay (Herskowitz Lab) This assay is semiquantitative. Cells can even be recovered from underneath the overlaid agar. The method could be used for any enzyme with a colorimetric assay.
Pheromone Halo Assay(Dohlman Lab) This is a bio-assay that measures the responsiveness of cells to a factor pheromone. The assay is easy to conduct and the results are usually unambiguous and highly reproducible.
Liquid ß-gal Assays (Dohlman Lab) This protocol is used to quantitatively measure the pheromone response over a range of pheromone concentrations.
ß-gal Lift Protocol (Dohlman Lab) This method is used to screen for and examine mutants with altered pheromone responses.
Protocols for shuttle mutagenesis/epitope-tagging of a yeast gene with mTn-lacZ/LEU2 (Yale Yeast Analysis Center) mTn-lacZ/leu2 can easily be inserted at mutiple sites in a given gene. The mutagenized DNA is then transformed into yeast, where it replaces the chromosomal locus by homologous recombination. The transposon insertions create a pool of insertion/disruption alleles. Insertions that generate in-frame fusion of the coding region to lacZ can be used to monitor and quantify gene expression, via assays for beta-gal activity. The fusion protein can also be immunodetected using antibodies directed against beta-gal.
Confirmation PCR (Saccharomyces Genome Deletion Project) To study the genetics of yeast, yeast gene ORF is deleted by PCR based strategy to generate mutated strain. Confirmation PCR is used to verify if the yeast ORF deletion is successful
