title: RussellDG-Mtb-Rv+CDC1551 spotted oligo array
status: Public on Nov 14 2007
contact.city: Ithaca
contact.first: Kyle
contact.middle: Herman
contact.last: Rohde
contact.email: khr4@cornell.edu
contact.state: NY
contact.address: Vet. Med. Center, C5 101
contact.department: Microbiology and Immunology
contact.country: USA
contact.institute: Cornell University
contact.phone: 607-253-4017
organism: Mycobacterium tuberculosis
manufacture_protocol: The version 1.1 Mtb 70-mer oligo library was used.  It contains a totall of 4320 70-mers, including targets for 3,924 ORFs annotated for strain H37Rv by Sanger plus an additional 371 ORFs from strain CDC1551 (TIGER) with less than 97% homology to H37Rv.  Oligos are Tm normalized to 79C +/- 5 degrees C.  Negative controls are 25 randomized 70-mers.  Positive control - mix of 4295 Mtb specific 70-mers.  Oligos were spotted using a Genomic Solutions (previously BioRobotics), MicroGrid Pro arrayer equipped with Micro-Spot 2500 pins (BioRobotics) onto Corning UltraGap slides. Each target wass spotted in duplicate  After spotting slides were UV irradiated at 200 milli-Joules in a BioRad UV box and then baked  for 1-2 hours at 85C.  They were then washed with 1%SDS for 2 minutes, then 2X Milli-Q water wash, dunk in 100% ethanol for 10 seconds and dried by centrifugation at 600RPM for 3 minutes.