title: IGBMC-Mouse-10K-MM11E
status: Public on Jun 30 2012
contact.city: Illkirch
contact.first: Bernard
contact.last: Jost
contact.address: 1, rue  Laurent Fries
contact.country: France
contact.institute: IGBMC
organism: Mus musculus
manufacture_protocol: 1. Microarrays contain 10,752 murine cDNA clones obtained from five different cDNA bank sources, corresponding to 2150 genes.
  2. Plasmid templates for the 10,752 clones were isolated from bacterial clones and inserts were amplified by PCR.
  3. PCR products were checked on agarose gels and purified by filtration on 96-well MAFB NOB50 multiscreen-FB filter plates (Millipore, Bedford, MA, USA)
  4. Purified PCR products were dried by evaporation at room temperature in a speedvac® (SC210A, Savant Instruments, Holbrook, NY, USA)
  5. Dried PCR products were resuspended in 75 % formamide/25% milliQ water at a final concentration of 0.5 µg/µl and were incubated for 48 h at 4C.
  6. The products were transferred to a 1536 well plate (#3950, Corning, NY, USA).
  7. A Microdgrid II arrayer (Genomics Solutions, Cambridge, UK) was used to print each product on ultraGAPS Coated Slides (# 40016, Corning Ltd) at 20-22C, in 45% relative humidity.
  8. Arrays were dried 48 h in a dessicator, then DNA was cross-linked to the slide by UV irradiation (600 mJoules) in a UV2500 Stratalinker (Stratagene, La Jolla, CA, USA).