title: E. coli K12, Sakai and EDL933 genes (MGW oligonucleotide)
status: Public on Nov 13 2008
contact.city: Montreal
contact.first: Sébastien
contact.middle: P
contact.last: Faucher
contact.email: sebastien.faucher@umontreal.ca
contact.state: QUEBEC
contact.address: C.P. 6128 Succ. Centre-Ville
contact.department: Microbiology and Immunology
contact.country: Canada
contact.institute: University of Montreal
contact.phone: 514-343-6111 #3135
organism: Escherichia coli
manufacture_protocol: Oligo set used is the E. coli O157 set commercially provided by MWG-Biotech and supplemented with 104 additional oligos requested by Vic Gannon's group. All were 50'mers resuspended in fresh 50%DMSO to a final concentration of 50uM using a Biomek 2000 robotic liquid handler (Beckman Coulter Inc). Print plates were sealed and stored at -20C when not in use. Prior to printing arrays the oligo plates were allowed to come to room temperature then heated for 5min at 60C. Once cooled they were centrifuged for 3min at 3000rpm to collect all liquid prior to arraying. Arrays were printed on Corning UltraGAPS slides using Telechem SMP3 pins in a BioRad ChipWriter Pro arrayer. Arrays were printed at room temperature (21-23C) and at 58-60% relative humidity. Arrays were printed with 24 pins in a 4x6 arrangement such that each subgrid contains 18x16 spots with an interspot distance of 245 microns centre to centre and a spot diameter of 100 microns. Duplicate supergrids were created lengthwise across the slide so that two spots per oligo exist at separate locations on the array. When spotting was complete, the chamber humidification was turned off and the arrays were allowed to "dry" in the print chamber for a minimum of 6 hours. The slides were then cross-linked at 600mJ and packaged in the original slide holders that they were received in from Corning. For shipping, arrays were packaged with dessicant in foil heat sealed bags.