title: GBC_ FOAR03-S1-0002
status: Public on Mar 04 2008
contact.city: Vancouver
contact.first: Juergen
contact.last: Ehlting
contact.email: je@uvic.ca
contact.state: BC
contact.address: 321 - 2185 East Mall
contact.department: Michael Smith Laboratories
contact.country: Canada
contact.institute: University of British Columbia
organism: Arabidopsis thaliana
manufacture_protocol: The Arabidopsis Genome Oligo Set Version 1.0 (Operon Biotechnologies, Huntsville, AL, USA) consists of 26090 70mer oligonucleotides (http://operon.com/arrays/oligosets_arabidopsis.php).  As positive controls, we included oligos for twelve housekeeping genes (Operon). As negative controls, we synthesized four oligonucleotides specific for human genes with no similarity to any Arabidopsis gene and included twelve oligonucleotides with no similarity to any Arabidopsis gene (Operon). We used a PCR amplified green fluorescent protein (GFP) cDNA (Invitrogen, Carlsbad, CA, USA) as an orientation marker. Oligonucleotides were resuspended in 384 well flat bottom plates (Nunc, Rochester, NY, USA) to a concentration of 100 mM in 3 x SSC. Oligos were printed on MicroGrid II robots using Microspot 10k pins (Biorobotics, Huntington, UK) depositing approximately 0.5 nl (0.0075 pmole) of each oligo onto EZ rays aminosilane slides (Apogent Discoveries, Hudson, NH, USA). The pitch of the grid used for this library was 0.2 mm. Oligos were UV cross linked at 3000 x 100 μJ using a UV Stratalinker 2400 (Stratagene, La Jolla, CA, USA).  We spotted single spots for the 26090 target oligonucleotides; in addition, each subgrid contained six replicate spots of each of the four human negative controls, three spots of equally distributed Operon negative controls, a single spot of each of the twelve house keeping controls, and GFP marker on each corner of the subgrid. The location of each oligo is given in this platform file.