title: Murine 22K cDNA array version 38.1
status: Public on Aug 29 2007
contact.city: Vienna
contact.first: J. Andrew
contact.last: Pospisilik
contact.email: pospisilik@imp.univie.ac.at
contact.address: Dr. Bohr-gasse 3
contact.country: Austria
contact.institute: IMBA
organism: Mus musculus
manufacture_protocol: Arraying
  1. Transfer PCR reactions to 96-well V-bottom tissue culture plates (Costar).Add 1/10 vol. 3M sodium acetate (pH 5.2) and equal volume isopropanol. Store at -20 C for a 2 hours.
  2. Centrifuge in Sorvall at 3500 RPM for 45 min. Rinse with 70% EtOH, centrifuge again and dry.
  2. Resuspend DNA in 12ul 3X SSC for a 4 hours and transfer to flexible U-bottom printing plates.
  4. Spot DNA onto poly-l-lysine slides with an arrayer.
  Post processing.
  1. Rehydrate arrays by suspending slides over a dish of warm ddH2O. (~1 minute)
  2. Snap-dry each array (DNA side up) on a 100°C hot plate for 3 seconds.
  3. UV X-link DNA to the glass by using a Stratalinker set for 60 milliJoules
  4. Dissolve 5g of succinic anhydride (Aldrich) in 315mL of n-methyl-pyrrolidinone.
  5. To this, add 35mL of 0.2M NaBorate pH 8.0 (made by dissolving boric acid in water and adjusting the pH with NaOH), and stir until dissolved.
  6. Soak arrays in this solution for 15 minutes with shaking.
  7. Transfer arrays to 95°C water bath for 2 minutes
  8. Quickly transfer arrays to 95% EtOH for 1 minute.
  9. Remove excess liquid from slides by spinning the rack of slides on microtiter plate carriers at 500rpm.