title: University of Tasmania Listeria monocytogenes 3K
status: Public on Jun 07 2007
contact.city: Hobart
contact.first: John
contact.middle: Phillip
contact.last: Bowman
contact.email: john.bowman@utas.edu.au
contact.state: Tasmania
contact.address: College Road, Sandy Bay
contact.department: School of Agricultural Science
contact.country: Australia
contact.institute: University of Tasmania
contact.phone: 61 03 6226 6380
organism: Listeria monocytogenes
description: Custom oligo set based on Listeria monocytogenes EGD-e (cat. no. 812002) and control probe set (cat. no. 812099). Oligos were arrayed with quill pens at the Australian Genome Research Facility, Inc., Walter & Eliza Hall Medical Institute, Level 7, 1G Royal Parade, Parkville, Melbourne, Victoria, 3050, Australia
manufacture_protocol: 70 bp DNA oligonucleotides-based spotted array on glass slides. For full details of methodology contact Dr. Stephen Wilcox;
  stephen.wilcox@agrf.org.au; Australian Genome Research Facility, Inc., Walter & Eliza Hall Medical Institute, Level 7, 1G Royal Parade, Parkville, Melbourne, Victoria, 3050, Australia.
  RNA conversion to cDNA and postlabelling with Cy3 and Cy5 was performed with the SuperScript Indirect cDNA Labeling System (Invitrogen).
  Prehybridization was performed in hybrization solution (25% vol./vol. formamide, 5 X SSC buffer – 0.75 M NaCl, 0.075 M trisodium citrate, pH 7.0, 0.1% sodium dodecyl sulfate) containing 10 mg ml-1 bovine serum albumin for 45 min at 42°C. Slides were then rinsed twice in distilled water and air-dryed. Hybridisation was performed in a humid hybridisation chamber at 42°C for 16-20 h using a hybridisation solution containing 0.42 ug ul-1 human Cot1 DNA, 0.62 ug ul-1 polyA and 0.83 ug ul-1 salmon sperm DNA. Slides were washed at room temperature in 1 X SSC buffer containing 0.2% SDS for 5 min and again in 0.1 X SSC buffer containing 0.2% SDS for 5 min. The slides were then further washed twice in 0.1 X SSC buffer at room temperature for 2 min.  Slides were then dried and subsequently scanned using a GenePix 4000A scanner (Axon Instruments).