title: ICAT_UMBPB Pyrus communis 3.5K cDNA microarray
status: Public on Jul 02 2011
contact.city: Lisbon
contact.first: Vanessa
contact.middle: Tolentino
contact.last: Pereira
contact.email: vanessaisadora@yahoo.com
contact.address: Ed. ICAT, Campus da FCUL, Campo Grande
contact.department: Unit of Molecular Biology and Plant Biotechnology
contact.country: Portugal
contact.institute: FCUL/ICAT/BioFIG
contact.phone: 00351217500163
organism: Pyrus communis
manufacture_protocol: 1. Amplified  cDNA inserts were purified with MultiScreen96-PCR plates (Millipore), transferred to printing plates (Microtiter V plates, Sigma), dried through passive water evaporation in a protected environment and resuspended in 10 µl of Pronto! Universal Spotting Solution (Corning, Life Sciences, USA) to a final concentration of 150 to 200 ng/µl.
  2. cDNAs were spotted in duplicate onto glass UltraGAPS™ Coated Slides (Corning, Life Sciences, USA) using VersArray ChipWriter Compact System ® (Bio-Rad Lab., Hercules, CA, USA). Humidity: 50%-55%. Temperature: 22°C. Pins: 12 printing pins (Telechem International, Sunnyvale, CA, USA). Pin configuration: 2 x 6. Number of repeats: spots are repeated once on the same subgrid in the same order. Printing plates: 36 x 2 (96 wells). Printing scheme: 12 subgrids, each subgrid with 24 columns and 24 rows. Distance between spots: 300μm. This is a condensed version of the physical array, 7K. The full array linked as a supplementary file.
  3. After printing, microarrays were processed with heat and UV crosslinking following manufacturer's instructions (Corning, Life Sciences, USA).