title: DGC Drosophila v1 6k array
status: Public on Nov 17 2016
contact.city: Berlin
contact.first: Ansgar
contact.last: Klebes
contact.email: ansgar.klebes@web.de
contact.address: Chausseestr. 117
contact.department: Biology
contact.country: Germany
contact.institute: Humboldt University Berlin
contact.phone: +49-30-20938174
organism: Drosophila melanogaster
manufacture_protocol: Spotted cDNA microarrays were produced essentially as  described in Klebes et al., 2002 and DeRisi et al., 1997. In brief, the Drosophila Gene Collection  (DGC 1.0, kindly provided by the Berkeley Drosophila  Genome Project) of 5,849 nonredundant cDNAs was amplified  using universal primers essentially as described at Berkeley Drosophila Genome Project resources [http://www.fruitfly.org/about/methods/ampUniGene.html].   The amplification products were purified with 96-well  format Qiagen PCR purification columns. All PCR products  were analyzed on agarose gels, and reactions with no  detectable product, multiple bands or bands of unexpected  size were repeated using Herculase (Amersham) or Expand  polymerase (Roche). One hundred and seventy-five cDNAs  from our lab collection were amplified and added to the set,  resulting in a total of 6,024 cDNAs. Reactions were arrayed  into 384-well plates and printed on poly-L-lysine-coated  glass slides using a linear servo arrayer and ArrayMaker  Version 2 control software. To minimize background caused  by oxidation of the polylysine coating, slides were pretreated  by a protocol suggested by Paul Ebert. Before post-processing,  slides were incubated for 5 min in 3x SSC; 0.2% SDS at  65°C, washed first in water and then in 95% ethanol, followed by centrifugation to dry the slides. Slides were then treated, and the hybridization reaction was carried out as described at Drosophila Genome Project resources [http://www.fruitfly.org/about/methods/ampUniGene.html]. We indirectly labeled the hybridization probes by incorporation of amino-allyl modified nucleotides in a first-strand cDNA RT reaction. Monofunctional Cy5 or Cy3 dye (Amersham) was subsequently coupled to the reactive residues. Multiple hybridizations were carried out, and dye labeling was reversed to avoid systematic bias.