title: Ambion mirVana™ miRNA Probe Set 1564V2 Oct06_IZKF (condensed)
status: Public on Sep 10 2010
contact.city: Wuerzburg
contact.first: Burkhard
contact.middle: Gerhard
contact.last: Kneitz
contact.email: Kneitz_B@klinik.uni-wuerzburg.de
contact.address: Oberdürrbacher Str. 6
contact.department: Klinik und Poliklinik für Urologie und Kinderurologie
contact.country: Germany
contact.institute: University of Wuerzburg
contact.phone: +49-931-201-32700
organism: Homo sapiens
  Mus musculus
  Rattus norvegicus
description: support = Nexterion® Slide E, coating = epoxy
  
  The Platform data table reflects a condensed representation of the array's quadruplicate features.  The full array layout representing all of the individual features is linked as a supplementary file at the foot of this record.
manufacture_protocol: 1. Oligos are arrayed in 96 well plates. Each well contains 500 pmol of 42–46 nucleotides (nt) long oligos and consist of an 18–24 nt segment that targets a specific known human, mouse, or rat miRNA, plus spacer and attachment sequences for coupling to the surface of the microarray slide
  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
  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