title: HMS Macaca mulatta 2.7K V1.0
status: Public on Feb 27 2006
contact.city: Boston
contact.first: Andrei
contact.middle: V
contact.last: Tkatchenko
contact.state: MA
contact.address: 220 Longwood Avenue, B2-201
contact.department: Neurobiology
contact.country: USA
contact.institute: Harvard Medical School
organism: Macaca mulatta
manufacture_protocol: cDNA subtractions and construction of cDNA libraries were performed as previously described. Briefly, subtracted cDNA libraries were generated using the PCR-Select cDNA Subtraction Kit (BD
  Biosciences Clontech, Mountain View, CA) by cloning the secondary PCR
  products into the pCRII-TOPO vector (Invitrogen, Carlsbad, CA). Two subtractions were carried out resulting in two cDNA libraries. One library was
  enriched in clones potentially up-regulated in the retina of the closed eye and the other was enriched in cDNAs potentially down-regulated. One thousand
  clones from each library were organized in 384-well plates. Bacterial clones
  from these plates were manually printed onto nylon filters using a 384-pin tool
  (V&P Scientific, San Diego, CA) to prepare two identical sets of bacterial
  colony filters. These filters were hybridized with subtracted probes, and 535 differentially-expressed cDNAs were amplified by PCR, purified, dried and resuspended in 5xSSC. The cDNA library was then spotted onto glass slide using QArray2 microarrayer.