title: Rat 10K gene set
status: Public on Dec 23 2011
contact.city: Stillwater
contact.first: Tingting
contact.last: Weng
contact.email: tingting.weng@okstate.edu
contact.state: OK
contact.address: 264 McElroy Hall
contact.department: Physiological Sciences
contact.country: USA
contact.web_link: http://www.lungmicroarray.org
contact.institute: Oklahoma State University
contact.phone: 4057443348
organism: Rattus norvegicus
manufacture_protocol: 1.  The Pan Rat 10K Oligonucleotide Set (MWG Biotech Inc., High Point, NC, USA) contains aminated 50-mer oligonucleotides.
  2. Oligos (25 µM) were amino-modified and resuspended in 3 x SSC printing buffer
  3. The oligos represent 6,221 known genes, 3,594 EST and 169 Arabidopsis negative controls.
  4. Each oligo was spotted in triplicate on three identical 18 x 18 mm blocks in one slide.  The spot-spot distance was 160 µm and the space between blocks was 4 mm.  Each block consists of 16 subarrays and can be used as an individual slide or replicate.
  5.The oligos were arrayed from 26 384-well microplates onto Epoxy glass slides using an OminiGrid 100 arrayer (GeneMachine, San Carlos, CA)
  6. To immobilize oligos, the slides were incubated for 2 days at room temperature and 50% humidity
  7. Slides were washed one time with 0.2% SDS for 2 min and then three times with ddH2O
  8. Finally, slides were incubated in ddH2O for 20 min at 50oC.  Printing spots were uniform as assessed by SYTO-6 staining.