title: University of Cape Town Cowpea 9K drought stress SSH library
status: Public on Mar 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: Vigna unguiculata
manufacture_protocol: 1. PCR fragments of a cloned cDNA SSH library are arrayed in Genetix X7022 384-well plates.
  2. Each well contains a final concentration of 125 ng/µl PCR product in 50% DMSO spotting solution (v/v).
  3. Between printing runs the plates are stored at -20°C.
  4. Prior to printing the plates are incubated at room temperature for 30 min to aid temperature equilibration.
  5. Plates are centrifuged at 2000 x g for 5 minutes prior to printing.
  6. Printing is performed on the Molecular Dynamics Generation III spotter.
  7. Seals are removed and plates placed in the spotter immediately prior to initiating the printing process to prevent unnecessary evaporation.
  8. A relative humidity of 50% is maintained in the spotter cabinet during printing runs.
  9. Following completion of the printing run the plates are sealed with a new Abgene adhesive plate seal and stored at -20°C.
  10. Each slide is made up of 2144 and 2016 probes from the F and R libraries, respectively, plus 448 probes which are control or blank spots. Each probe is printed twice to make a total of 9216 probes on each array.