title: Uni Wuerzburg Listeria monocytogenes 13.8K whole genome array
status: Public on May 01 2006
contact.city: Wuerzburg
contact.first: Biju Joseph
contact.last: Ampattu
contact.email: bjoseph@hygiene.uni-wuerzburg.de
contact.address: Josef-Schneider Str 2
contact.department: Department of Microbiology
contact.country: Germany
contact.institute: University of Wuerzburg
organism: Listeria monocytogenes
description: Epoxy coated slides used to spot 70mer oligonucleotides
  Whole genome DNA microarrays that contain synthetic 70mer oligodeoxyribonucleotides covering all ORFs of the Lm genome. The oligonucleotides (sequences available at http://www.operon.com/arrays/oligosets_listeria.php) were spotted on epoxy coated glass slides from Quantifoil according to manufacturer’s instructions by M. Frosch (Institut für Hygiene and Mikrobiologie, Würzburg, Germany). Each oligo was spotted four times on a slide to generate four replicates for each oligo on a slide.
manufacture_protocol: ARRAY PRINTING
  1. Dissolve oligonucleotide probe or PCR product in the appropriate spotting solution to obtain the
  recommended final probe concentration:
  DNA Probes Final Spotting Concentration
  Oligonucleotides 10 - 20 ìM
  PCR products 0.1 – 1 mg/ml
  2. Transfer an appropriate volume of probes to a microtiter plate.
  3. Set up the arrayer according to the manufacturer’s recommendations. If you were previously
  using slides that were thicker than 1.0 mm, for optimal spotting you may need to re-calibrate
  the distance between the slide surface and the spotting pins.
  4. Print the substrates at 40-50% relative humidity at 20 to 25°C.
  DNA IMMOBILIZATION
  1. Incubate printed microarray slides in humidity chamber at room temperature for 30 min (see
  appendix for details of how to prepare this chamber) to ensure quantitative immobilization.
  2. Proceed to washing