title: University of Cape Town Arabidopsis thaliana 7200 cDNA array
status: Public on Nov 22 2010
contact.city: Pretoria
contact.first: Dave
contact.middle: K
contact.last: Berger
contact.state: Gauteng
contact.address: Room 6-26, Agricultural Sciences Building, University of Pretoria, Lunnon Rd
contact.department: Plant Science, Forestry & Agricultural Biotech. Instit. (FABI)
contact.country: South Africa
contact.web_link: http://fabinet.up.ac.za/mppi/
contact.institute: University of Pretoria
contact.phone: +27-12-4204634
organism: Arabidopsis thaliana
manufacture_protocol: 1. Mendel Biotechnology L35 collection of Arabidopsis ecotype Col-0 cDNA library were amplified using vector specific primers (5'CGCTCTAGAGGATCCAAGCTTACGT-3' and 5'ACCGGTCCGGAATTCCCGGGTCGAC-3')
  2. PCR fragments were purified using Multiscreen PCR purification plates
  3. Purified PCR fragments were arrayed in Genetix X7022 384-well plates.
  4. Each well contained a final concentration of 125 ng/µl PCR product in 50% DMSO spotting solution (v/v).
  5. Between printing runs the plates were stored at -20°C.
  6. Prior to printing the plates were incubated at room temperature for 30 min to aid temperature equilibration.
  7. Plates were centrifuged at 2000 x g for 5 minutes prior to printing.
  8. Printing was performed on the Molecular Dynamics Generation III spotter.
  9. Seals were removed and plates placed in the spotter immediately prior to initiating the printing process to prevent unnecessary evaporation.
  10. A relative humidity of 50% was maintained in the spotter cabinet during printing runs.
  11. Following completion of the printing run the plates were sealed with a new Abgene adhesive plate seal and stored at -20°C.