title: Wolbachia pipientis wMel microarray
status: Public on May 11 2009
contact.city: Baltimore
contact.first: Julie
contact.middle: C
contact.last: Dunning Hotopp
contact.email: jdhotopp@som.umaryland.edu
contact.state: MD
contact.address: 20 Penn St.; HSF-II; S203B
contact.department: Institute for Genomic Sciences
contact.country: USA
contact.institute: Univeristy of Maryland Baltimore
contact.phone: 410-706-5673
organism: Wolbachia pipientis
manufacture_protocol: A wMel microarray was constructed which represents 95% of the unique ORFs. Primers were designed from the unique predicted genes annotated in wMel. In order to obtain pure amplicons devoid of background amplification of host genes, all amplicons were produced, where possible, from two separate clones from the wMel genome sequencing project. Genes in physical gaps where a sequencing clone was not available were amplified directly from genomic DNA from D. melanogaster infected with wMel. Amplicons (50-1000 bp) were produced using AmpliTaq (Applied Biosystems), purified using filtration plates (Millipore), and analyzed on agarose gels. The purified amplicons were diluted 1:1 with DMSO, spotted in quadruplicate onto Corning UltraGap slides with a Lucidea printing robot (GE Healthcare), and irradiated with ultraviolet light.