1.Oligos are arrayed in Genetix 384 Well Microarray Plates (Cat# X7020). Each well contains 600 pmol of 70-mer oligo. 2. Resuspend oligos in 17 µL of Telechem Microspotting solution 1X. 3. Seal plates with Corning seals and after oligos resuspenction print probe with the MicroGrid II Biorobotics spotter. 4. Print is performed with 48 Stealth pins. 5. After printing run plates are stored at -20 C degree. 6. Every 4 months plates are lyophilized and resuspended in adjusted water volume according to the number of spotting runs. This, to maintain the same volume and concentration in all plate well.
This set consists of 21,329 70mer oligonucleotides (Operon version 2.0) designed on Human Unigene clusters, mainly in the 3 end terminal region. Each oligonucleotide is spotted by our robotic station Biorobotics Microgrid II, in one replicate on a MICROMAX glass slide SuperChip I (Cat No MPS696) provided by PerkinElmer Life Sciences Inc. The deposition is qualitatively assessed by a scan of the spotted slide at 10 micron resolution, using a GSI Lumonics scanner and a protocol that allows the detection of spotted features through the salts derived from the dried solution in which oligonucleotides are dissolved. Moreover all spotted batch are evaluated for the spot shape and oligo linking onto the slide surface using the vistra green method. Microarray slides that meet the quality standards, are washed as recommended by the supplier and the glass slides are stored, ready for use, in the dark under deep vacuum.