title: Listeria monocytogenes (EGDe) macroarray
status: Public on May 29 2009
contact.city: Porto
contact.first: Didier
contact.last: Cabanes
contact.email: didier@ibmc.up.pt
contact.state: Porto
contact.address: rua do Campo Alegre, 823
contact.country: Portugal
contact.institute: IBMC
organism: Listeria monocytogenes EGD-e
manufacture_protocol: The macroarrays were constructed by Milohanic et al (Mol Microbiol 2003. 47: 1613-1625). Whole-genome arrays were constructed by amplifying 500-bp-long PCR products specific for each gene using open reading frame (ORF)-specific primers. Ninety-nine per cent (2816 ORFs) of the 2853 predicted ORFs of the L. monocytogenes EGDe genome were amplified successfully.
  For macroarray preparation, nylon membranes were soaked in 10 mM TE, pH 7.6. Spot blots of ORF-specific PCR products and controls were printed using a Qpix robot. Immediately after spot deposition, membranes were neutralized for 15 min in 0.5 M NaOH, 1.5 M NaCl, washed three times with distilled water and stored wet at -20°C until use.