title: FhG_IGB_MBT Candida albicans oligo array V1
status: Public on Jul 15 2011
contact.city: Stuttgart
contact.first: Steffen
contact.last: Rupp
contact.address: Nobelstr. 12
contact.department: MBT
contact.country: Germany
contact.institute: Fraunhofer IGB
organism: Candida albicans SC5314
manufacture_protocol: 1. Oligos (Operon Biotechnologies Custom Array-Ready Longmers, Eurofins MWG Operon, Ebersberg, Germany) are stored at -20°C in Genetix plates (Order. Nr. X6003). Each well contains 600 pmol of 55-70-mer oligos.
  2. Allow plates to reach room temperature.
  3. Centrifuge oligo plates at 200 g for 2 min.
  4. Remove sealing foil and resuspend oligos in 12 µL 3*SSC/1.5 M Betain to 50 µM (working concentration 50 pmol/µL).
  5. Centrifuge oligo plates covered with plate lids at 200 g for 2 min.
  6. Place in appropriate location of biobank and start print run.
  7. Oligos are printed on epoxy coated glass slides (Schott, Jena, Germany) using the Microgrid II robot of Biorobotics (DigiLab Inc., Holliston, USA) by spotting 6,400 oligonucleotides in duplicate including 10 lambda oligonucleotide DNA positive controls. SMP3 pins (Arrayit Corporation, Sunnyvale, CA) are used for spotting.
  8. After the print run is completed, allow plates to dry through passive evaporation in a protected environment.
  9. For each subsequent preparation of these plates for a print run, add 12 µL water to the wells instead of 3*SSC/1.5 M Betain.
  10. After the spotting process, probes were immobilized on glass slides by incubation at room temperature for 30 min at high humidity (> 80 %) and adjacent drying at 80°C for 120 min. DNA-microarrays were stored under argon atmosphere in an exsiccator until used.
  11. For data analysis, a gal file was produced using the TAS Application Suite software version 2.7.1.18 from Genomic Solutions.