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      Genome-wide expression monitoring in Saccharomyces cerevisiae.

      Nature biotechnology
      Cell Division, genetics, Gene Expression Regulation, Fungal, Genome, Fungal, Light, Nucleic Acid Hybridization, methods, Oligonucleotides, chemical synthesis, Polymerase Chain Reaction, RNA, Fungal, analysis, RNA, Messenger, Reproducibility of Results, Saccharomyces cerevisiae, cytology

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          Abstract

          The genomic sequence of the budding yeast Saccharomyces cerevisiae has been used to design and synthesize high-density oligonucleotide arrays for monitoring the expression levels of nearly all yeast genes. This direct and highly parallel approach involves the hybridization of total mRNA populations to a set of four arrays that contain a total of more than 260,000 specifically chosen oligonucleotides synthesized in situ using light-directed combinatorial chemistry. The measurements are quantitative, sensitive, specific, and reproducible. Expression levels ranging from less than 0.1 copies to several hundred copies per cell have been measured for cells grown in rich and minimal media. Nearly 90% of all yeast mRNAs are observed to be present under both conditions, with approximately 50% present at levels between 0.1 and 1 copy per cell. Many of the genes observed to be differentially expressed under these conditions are expected, but large differences are also observed for many previously uncharacterized genes.

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          A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae.

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            Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction

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              Light-directed, spatially addressable parallel chemical synthesis

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