title: Operon Oligonucleotide Set Innate Immunity and Aspergillosis 7MM
status: Public on Apr 17 2012
contact.city: Wuerzburg
contact.first: Susanne
contact.middle: Elma
contact.last: Kneitz
contact.email: susanne.kneitz@uni-wuerzburg.de
contact.address: Biozentrum, Am Hubland
contact.department: Physiologigcal Chemistry
contact.country: Germany
contact.institute: University of Wuerzburg
contact.phone: +49-931-31 86526
organism: Homo sapiens
description: Nexterion® Slide E
manufacture_protocol: 1. Oligos are arrayed in 96 well plates. Each well contains 500 pmol of 55–70 nucleotides (nt) long oligos and consist of an 18–24 nt segment that targets  specific known human cytokine, chemokine and innate immunity genes
  2. Add  3 x SSC + 1,5 M Betaine (in Rnase-free water) to 10 uM
  3. Seal plates and wrap them in saran wrap
  4. Let it resuspend in a humid chamber on a shaker (60 rpm) over night
  5. Centrifuge plates at 2000 rpm for 3 minutes.
  6. Rearray 10 µL into 384-well, V-bottom plates
  7. Centrifuge plates at 2000 rpm for 3 minutes.
  8. Remove seals and cover with plate lids. Place in appropriate location of plate cassette. This should be done with first plates just before print run is started to minimize evaporation time before printing. For second and third cassettes, wait until 15 minutes before next cassette is needed to begin centrifugation.
  9. After the print run is completed, allow plates to dry through passive evaporation in a protected environment.
  10. Spots are printed in quadruplicates, assiciated data contain median values of quadruplicates
  11. For each subsequent preparation of these plates for a print run, add water to the wells instead of print buffer. The amount of water should be decreased by 0.25 µL per print run, as this is the amount drawn up by the pin capillary during each dip.