title: Interference of components of the PEP:phosphotransferase system (PTS) with PrfA of L. monocytogenes
status: Public on Feb 01 2007
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
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