title: Sunflower organ-specific medium-scale cDNA array
status: Public on Jan 29 2008
contact.city: Castelar
contact.first: Paula
contact.last: Fernandez
contact.email: pfernandez@cnia.inta.gov.ar
contact.state: Buenos Aires
contact.address: N. Repetto y Los Reseros S/N
contact.country: Argentina
contact.institute: INTA Castelar
organism: Helianthus annuus
manufacture_protocol: Five ul from each purified PCR cDNA products (317 in total) were transferred into 384 well plate containing 5 ul of DMSO 100%,spotted onto coated glass-slides in the working solution plates and  printed onto UltraGap Slides (Corning) using a Versarray chipwriter (Bio-Rad) with Stealth Micro Spotting quill pins (Telechem). Humidity was set at 45%, and temperature was 22-25°C. Pins were loaded in the working plate during 0.5 sec. Distance between spots was 400 um, Dwell time for printing was 0.05 sec. Four replicas from each cDNA were spotted per slide. Pins were loaded in the working plate and before printing the slides, a blotting slide was printed 5 times in order to discard the excess of cDNA on the pins. After printing on slides, and before loading a new cDNA, pins were washed with 3 cycles of 5 sec. wash with water, followed by 1 sec vacuum and a final 10 sec vacuum time . After printing was completed, cDNA were UV crosslinked to the slides on a CL-1000 UV crosslinker (UVP). Printed slides were stored in a dissecator.