title: USouthAL_Rickettsia prowazekii Madrid E 1.5K array version 1
status: Public on Aug 06 2008
contact.city: Mobile
contact.first: Jonathon
contact.last: Audia
contact.email: jaudia@jaguar1.usouthal.edu
contact.state: AL
contact.address: 307 University Blvd N
contact.department: Microbiology and Immunology
contact.country: USA
contact.institute: University of South Alabama
contact.phone: 251-460-6929
organism: Rickettsia prowazekii str. Madrid E
manufacture_protocol: Glass slides were spotted with (where possible) two unique 70-mer oligonucleotides corresponding to all 835 annotated ORFs from the R. prowazekii genome sequence.  70-mer oligonucleotides were designed from these sequences by using ArrayOligoSelector.  All 70-mer oligonucleotides were verified as specific for R. prowazekii and not Mus musculus (the host cell used for rickettsial infections) under the conditions of hybridization used in this study using the BLAST algorithm.  The arrays consisted of two unique 70-mer oligonucleotides for 666 ORFs and a single unique oligonucleotide for 169 ORFs.  The oligos were synthesized by using standard methods by illumina (San Diego). The oligonucleotides were dissolved at a concentration of 50 mM in 3 × SSC with 0.75 M betaine and were printed in duplicate on MWG Epoxy slides (MWG Biotech, Ebersburg, Germany) by a locally constructed linear servo arrayer (after the DeRisi model).  The oligonucleotides were spotted in triplicate.  In addition, oligonucleotide probes corresponding to 32 mouse genes were included as negative controls and oligonucleotide probes corresponding to 10 Arabidopsis genes (SpotReport® Oligo Array Validation System, Stratagene) were included as cDNA synthesis validation controls.