title: UMBI_Drosophila_1440_1
status: Public on Feb 10 2007
contact.city: College Park
contact.first: Subhamoy
contact.last: Pal
contact.state: MD
contact.address: 5126 Plant Sciences Building
contact.department: Center for Biosystems Research
contact.country: USA
contact.web_link: http://www.umbi.umd.edu
contact.institute: University of Maryland
organism: Drosophila melanogaster
description: Spotted oligonucleotide microarray with a high representation of genes important for Drosophila immune responses.
manufacture_protocol: 464 Genes important for Drosophila immune responses were selected from a literature survey and previous experiments. Primers were designed to be between 200 and 600 base pairs using manual techniques and Primer3.  These PCR primers were used to amplify the selected oligonucleotide regions from Genomic DNA, and run on agarose gels to confirm amplification success and specificity.  The reaction mixture to produce each amplicon contained 50 ng Drosophila genomic DNA, 1 uM forward primer, 1 uM reverse primer, 1xTitanium Taq (Invitrogen) and 0.5 mM dNTP. The following PCR protocol was used: an initial 95C denaturation step for 5 minutes followed by 20 cycles of 30 seconds of 95C denaturation, 30 seconds of 60C annealing, and 45 seconds of 75C extension. They were purified using QIAquick 96 PCR Purification Kit according to manufacturer's protocol and suspended in 3XSSC.
  The purified DNA was spotted onto Corning GAPSII coated glass slides using a Affymetrix Arrayer 417, at the CBR/UMBI Core Microarray Facility.