title: MERH03-S1-0003 (Rhodococcus sp. RHA1 11K)
status: Public on Apr 09 2008
contact.city: Vancouver
contact.first: Shawn
contact.last: Anderson
contact.email: shawn.anderson@vch.ca
contact.state: B.C.
contact.address: The Prostate Centre at Vancouver General Hospital, 2660 Oak Street
contact.department: Urological Sciences
contact.country: Canada
contact.institute: University of British Columbia
contact.phone: 604-875-4111 (63436)
organism: Rhodococcus jostii RHA1
manufacture_protocol: Oligonucleotides are consolidated into 384-well plates, lyophilized by speed-Vac, and resuspended in 3x SSC to a printing concentration of 30uM. Oligos are printed on aminosilane slides (Erie, Hudson, NH, USA) with a Biorobotics Microgrid II microarray printer (Genomic Solutions). Microspot 10K quill pins (Biorobotics) in a 48-pin tool were used to deposit <0.5 nL per spot onto the slide. The resulting microarrays have a 4 x12 subgrid layout with 392 spots per subgrid, each spot having approximate diameter and pitch of 90 µm and 190µm, respectively. A 280-bp GFP (green fluorescent protein) oligo was printed in subgrid corners to assist in placing on the grid. The slides were crosslinked in a UV Stratalinker 2400 (Stratagene) at 300mJ. Array quality was assessed by visual inspection and hybridization of representative slides from a print run by dye-labelled random 9-mer oligonucleotides. The quality control images were aquired via the ScanArray Express from Perkin Elmer at a 10 micron resolution and quantified with the Imagene 6.0 software suite.