title: UvA_MAD-mouse-22K-vs2.0
status: Public on Mar 09 2011
contact.city: Amsterdam
contact.first: Perry
contact.middle: D
contact.last: Moerland
contact.email: p.d.moerland@amsterdamumc.nl
contact.address: Meibergdreef 9
contact.department: Epidemiology and Data Science
contact.country: Netherlands
contact.institute: Amsterdam UMC
organism: Mus musculus
manufacture_protocol: 1.Mouse Oligo libraries were bought from Sigma Genosys (Cat # MOULIBST & Cat # MOULIB384B)
  2. Resuspend oligos in  25 ul 3 x SSC/1.5 M betain to 20 uM
  3. Seal plates with Corning seals.
  4. Shake plates near maximum rotational speed on flat-bed shaker for 1 minute.
  5. Centrifuge plates at 2000 rpm for 3 minutes.
  6. Remove seals and place plate in appropriate location of a Lucidea MicroArrayer (Amersham Pharmacia BioSciences,Piscataway,Nj,USA)
  7. Spot library on ultraGAPS slides (Corning)
  8. After the print run is completed, allow plates to dry through passive evaporation in a protected environment.
  9. Fix oligonucleotides by a heat treatment at 100C for a few seconds. For each subsequent preparation of these plates for a print run, add water to the wells instead of sodium phosphate 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.
  10. Complete fixation with a UV treatment at 150 mjoule
  11. Store arrays at dry and controlled location