title: Eurogentec C060C Yeast ORFs
status: Public on Jul 06 2006
contact.city: Brussels
contact.first: Bart
contact.last: Scherens
contact.address: Av. E. Gryson, 1
contact.department: Microbiology
contact.country: Belgium
contact.institute: IRMW-ULB
organism: Saccharomyces cerevisiae
description: The yeast 5803 ORFs MicroArray is composed of 32 subgrids, which follow a pattern of 8 metarows and 4 metacolumns. Each subgrid contains 20 columns and 20 rows with a dot spacing of 200 µm. Probes are spotted as closed duplicates. In order to provide a friendly environment, the first subgrid row on each array metarow contains all the normalization controls, which include the controls with serial dilution of the Renilla LuxA cDNA PCR product. In addition to this control, four serial dilutions of the sonicated yeast S288c genomic DNA are also located in the corner subgrids of the MicroArray.
manufacture_protocol: Comparative genome analysis of 6213 predicted Saccharomyces cerevisiae open reading frame (ORF) products with 13 yeast genome sequence related species proposed the elimination of 612 predicted ORFs that are not real genes (A. Malpertuy et al, FEBS Letters, Volume 487, 22 December 2000). The study identified the presence of 50 new genes.
  Specific pair of primers was designed by Pasteur Institute, Paris to amplify all 5803 ORFs non-spurious coding sequences. The primers were selected carefully to amplify only a specific region of each ORF.
  Universal sequences of 15 bases were incorporated on the 5' of each specific forward and reverse primer.
  The two-step PCR method has been employed to generate 5' amino-modified PCR for covalent attachment to the aldehyde-coated support.
  All the amplicons have an average length of 300 bp and they were controlled systematically by electrophoresis on agarose gels.