title: UWBMC Armigeres subalbatus 18K v1.0
status: Public on Apr 01 2010
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: Armigeres subalbatus
manufacture_protocol: Six cDNA libraries from adult female mosquitoes were used to generate 8,020 Expressed Sequence Tag (EST) clusters (Mayhew et al. 2007) from which 60-mer oligonucleotides were synthesized that represent 6,143 unique EST clusters.  Oligos were designed using OligoArray v 2.1, and these 6,143 oligonucleotides were used to construct our Armigeres microarray.  Ar. subalbatus microarray slides (Corning Ultragaps) were printed using a Genomic Solutions GeneMachines Omnigrid arrayer and SMP3 pins from TeleChem International (23˚C and 50% humidity) following established printing protocols.  Sixty-mer oligonucleotides were re-suspended at 40 μM in Pronto spotting buffer (Promega, Madison, WI).  Each microarray (19,200 total tool spots) was printed using 16 pins, resulting in 48 subgrids (20 x 20 at 0.215 mm pitch).  Features were printed in triplicate across the slide.
  
  Cytochrome C oxidase was printed at all four corners of each of the 48 subgrids and served as a positive control.  Negative controls on the array included blanks (spotting buffer with no oligo) and two mouse oligos, Mus 1 and Mus 2.  Blanks were interspersed within all subgrids, and Mus 1 and 2 were printed in triplicate like all other features.  A scanning control, Mus 3, was added because of its ability to be mixed with both Cy3 and Cy5 reactions, resulting in a 1:1 ratio on the array.  Mus 3 RNA (2.5 ng) was added as a spike during cDNA synthesis.  Forty-eight spots appeared as empty, i.e. a spot touched down on the slide but visited an empty well for its source visit.  These allow for new features to be added to the array without changing the general design.  Overall, the array had 6,143 unique features printed in triplicate (= 18,429 spots, including Mus 1 and 2), 192 positive control spots, 48 Mus 3 spots printed once per subgrid, and 495 blank spots.