title: Aedes aegypti hemocyte II
status: Public on Oct 17 2006
contact.city: Madison
contact.first: Bruce
contact.middle: M
contact.last: Christensen
contact.email: christensen@svm.vetmed.wisc.edu, mayhew@svm.vetmed.wisc.edu
contact.state: WI
contact.address: 1656 Linden Drive
contact.department: Pathobiological Sciences
contact.country: USA
contact.institute: University of Wisconsin-Madison
organism: Aedes aegypti
description: For the purpose of technical replication and to control for variation in hybridization across the slide, slides are designed such that each feature is desposited twice per grid, and that grid is duplicated on another part of the slide.  The result is that each feature is represented 4 times.
manufacture_protocol: 60-mer oligonucleotides were designed (for EST clusters generated for immune-response activated hemocytes from adult Ae. aegypti females (described in PMID: 15213157)) using OligoArray software version 1 (see PMID: 11934750).  Oligos were synthesized by Qiagen/Operon and resuspended to a final concentration of 40 uM in Pronto! Universal Spotting Solution (Corning Life Sciences).  The Biorobotics MicroGrid II microarrayer (Apogent Discoveries), fitted with 10K Tungsten pins was used with the following parameters: 0.275 pitch, -0.360 mm target height, 4mm/sec to 2mm/sec at 1mm above the slide deceleration, to acheive spots that are between 100 and 150 um in size.  The accompanying software TAS Application Suite version 2.2.2.6 was used to program the microarrayer.  Corning UltraGAPS slides (Cornign Life Sciences) were used as the array substrate.  After spotting, slides were subjected to 48 hours of vacuum desiccation at room temperature, then UV cross-linked at 600 mJ in a UV Stratalinker (Stratagene).