title: Technical Microbiology-Viable beer spoilage bacteria microarray V2
status: Public on May 23 2007
contact.city: Bochum
contact.first: Daniel
contact.middle: Gilbert
contact.last: Weber
contact.address: Bürkle-de-la-Camp Platz 1
contact.department: Molekulare Medizin
contact.country: Germany
contact.institute: IPA
organism: Pectinatus frisingensis
  Levilactobacillus brevis
  Lacticaseibacillus casei
  Loigolactobacillus coryniformis
  Megasphaera cerevisiae
  Pediococcus damnosus
  Pectinatus cerevisiiphilus
  Schleiferilactobacillus perolens
  Pediococcus inopinatus
description: Each oligonucleotide species-specific probe was spotted in 70 replicate spots on the microarray. Nine single beer spoilage bacteria species were detected with the developed prototype oligonucleotide microarray and also three mixes of two different beer spoilage bacteria species.
manufacture_protocol: Standard glass microscope slides were coated with a poly-L-lysine solution and stored for several weeks to allow the surface to become sufficiently hydrophobic. Species-specific probes were spotted in 3x SSC onto the glass surface using a robotic printing system. For routine experiments 50 μM was used as probe concentrations. The diameter of the spots was 150 μm and the space between the centres of two spots was 205 μm allowing the preparation of seven grids each containing 70 spot replicates. To distribute the oligonucleotides more evenly, the spots were rehydrated using a 1x SSC bath at 40°C until the spots were glistening, followed by snapped-drying at 120°C using a heat block. To increase the amount of hybridisable oligonucleotides, stably attached to each spot, the nucleic acids were cross-linked by UV irradiation.