title: Rhodococcus sp. RHA1 13.8K MERH03-S1-0004
status: Public on Nov 14 2008
contact.city: Houston
contact.first: Jacques
contact.last: Mathieu
contact.email: mathieu@rice.edu
contact.state: TX
contact.address: 6100 Main St. MS-317
contact.department: Civil and Environmental Engineering
contact.country: USA
contact.institute: Rice University
contact.phone: 703-307-5802
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.