TRANSCRIPTIONAL PROFILING ON ALL OPEN READING FRAMES OF SACCHAROMYCES-CEREVISIAE

Open reading frames (6116) of the budding yeast Saccharomyces cerevisi ae were PCR-amplified from genomic DNA using 12,232 primers specific t o the ends of the coding sequences; the success rate of amplification was 97%. PCR-products were made accessible to hybridization by being a rrayed at very high density on solid support media using various robot ic devices. Probes made from total RNA preparations were hybridized fo r the analysis of the transcriptional activity of yeast under various growth conditions and of different strains. Experimental factors that proved critical to the performance, such as different RNA isolation pr ocedures and the assessment of hybridization results, for example, wer e investigated in detail. Various software tools were developed that p ermit convenient handling and sound analysis of the large data quantit ies obtained from transcriptional profiling studies. Comprehensive arr ays are being distributed within the European Yeast Functional Analysi s Network (EUROFAN) and beyond. (C) 1998 John Wiley & Sons, Ltd.