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Application Note: ANCCSAA DNA Microarray Analysis of the Heat Shock Transcriptome of the Obligate Intracytoplasmic Pathogen Rickettsia prowazekii Jonathon P. Audia, Mary C. Patton, and Herbert H. Winkler Research Objective Key Words • Microarray • Transcriptome • Obligate Intracytoplasmic • Rickettsia prowazekii • R. prowazekii • oligonucleotide microarrays • Trypsinization • pathogenic bacteria • DNA Microarray • Obligate Intracytoplasmic • Assay • AssayAssure • swabs Identifying gene expression in Rickettsia prowazekii from mouse fibroblast cells. Isolating R. prowazekii total RNA from contaminating host cell nucleic acids for microarray analysis of all 835 putative R. prowazekii ORFs. Experimental Design Cells infected with R. prowasekii were harvested by trypsin treatment and collection by centrifugation. Testing Platform DNA oligonucleotide microarrays to perform highthroughput analysis of global gene expressions. AssayAssure Use Trypsinization and all subsequent steps were performed in 20% AssayAssure™ solution to preserve nucleic acid integrity. Thermo Scientific AssayAssure Sample Stabilizing Collection and Transport Devices Outcome An efficient methodology for the isolation of highly purified RNA from pathogenic bacteria like R. prowazekii using AssayAssure was developed. Reference Jonathon P. Audia, Mary C. Patton, and Herbert H. Winkler, DNA Microarray Analysis of the Heat Shock Transcriptome of the Obligate Intracytoplasmic Pathogen Rickettsia prowazekii, APPLIED AND ENVIRONMENTAL MICROBIOLOGY (2008) 74: 7809–7812 www.thermoscientific.com/assayassure © 2011 Thermo Fisher Scientific Inc. AssayAssure is a registered trademark of Sierra Molecular Corporation. All other trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries. Specifications,terms and pricing are subject to change. Not all products are available in all countries. Please consult your local sales representative for details. North America: USA/Canada +1 800 772 8871 ANCCSAA082011
