QUANTITATIVE PHENOTYPIC ANALYSIS OF YEAST DELETION MUTANTS USING A HIGHLY PARALLEL MOLECULAR BAR-CODING STRATEGY

A quantitative and highly parallel method for analysing deletion mutan ts has been developed to aid in determining the biological function of thousands of newly identified open reading frames (ORFs) in Saccharom yces cerevisiae. This approach uses a PCR targeting strategy to genera te large numbers of deletion strains. Each deletion strain is labelled with a unique 20-base tag sequence that can be detected by hybridizat ion to a high-density oligonucleotide array. The tags serve as unique identifiers (molecular bar codes) that allow analysis of large numbers of deletion strains simultaneously through selective growth condition s. Hybridization experiments show that the arrays are specific, sensit ive and quantitative. A pilot study with 11 known yeast genes suggests that the method can be extended to include all of the ORFs in the yea st genome, allowing whole genome analysis with a single selective grow th condition and a single hybridization.