title: PLBIO-DX - WH/GEX
status: Public on Mar 23 2012
contact.city: Mumbai
contact.first: Muralidhara
contact.last: Padigaru
contact.email: muralidhara.padigaru@piramal.com
contact.state: Maharashtra
contact.address: 1, Nirlon Compex, Off Western Express Highway
contact.department: Translational Research
contact.country: India
contact.institute: Piramal Lifesciences India Limited
contact.phone: +912230818802
organism: Homo sapiens
manufacture_protocol: A microarray chip with 37,632 transcripts representing the whole human genome were purchased from Operon Biotechnologies (Operon, USA). Every gene was confirmed for function and sequence with the latest RefSeq genome build and represented as a unique 70-mer oligonucleotides sequence. A 40μM concentration of the oligos was achieved by dissolving the oligos with 50% DMSO solution. The MTP plates were vortexed at 150 rpm for 2 hours and spun down at 1200 rpm in a swing out rotor centrifuge. Every gene/oligo was printed with a robotic arrayer – OG100 (Genomic Solutions, Ann Arbor, USA) on aminosilane coated glass slide (ArrayIT, USA). The relative humidity and the temperature were maintained at 45% RH and 23°C. The printing is achieved by contact disposition of 20nL of the oligo suspension onto the glass slide with stealth microarray pins (SMP3) having an uptake capacity of 250nL. Post printing, the slides were air dried for 6 hours in the printing environment and then cross-linked at 650 mL/sec2 with an UV cross-linker. The slides were then packed and stored under vacuum in the dark at room temperature.