Integrated RNA Amplification Micro Card

DESCRIPTION (provided by applicant): Nucleic acid analysis is the essential foundation of many biomedical research and diagnostic methods. Among these, microarray-based gene expression profiling has become a powerful research tool for understanding basic d isease biology, and a powerful diagnostic tool for profiling cellular states during disease progression and in response to treatment. However, preparation of RNA from biological samples for microarray analysis is presently a rate-limiting bottleneck. This is because RNA purification, amplification, and labeling remains a complex, time-consuming, labor-intensive, and expensive process. The primary goal of this proposal is produce devices and systems that dramatically reduce this burden, so that it is no long er an impediment to health-related research and diagnostics. A second goal is to increase experimental reliability, by reducing human error, thereby increasing the utility and efficiency of microarray expression analysis methods. To accomplish these goals Arcxis Biotechnolgies will develop a microamplification card (MAC), a small, integrated fluidic processing device, and a companion bench-top instrument that will ultimately completely automate sample preparation for microarray expression analysis. Multiple cards will run simultaneously in the MAC instrument, which will occupy only a small amount of laboratory bench space, greatly increasing laboratory productivity and decreasing utilization of laboratory resources. The initial goal of the proposed research is to demonstrate integration of linear mRNA amplification and amplified RNA purification on a microamplification card. The specific aims of the proposed research involve, first, fabrication of microamplification cards and actuating instrumentation, and se cond, systematic implementation and characterization of the biochemical reactions involved in linear RNA amplification and RNA purification. Both of these biochemical processes are well-established and highly characterized, allowing research to focus on th eir specific implementation in microamplification cards. Integration of biochemical processes of this complexity in a commercially viable format will constitute a significant achievement. First, it paves the way for rapid integration of additional processe s that will enable end-to-end processing, from cell lysates to pure, amplified, labeled mRNA ready for microarray analysis. Second, it will provide a platform fluidic technology that will find wider application in biomedical research in the future. Microar ray-based gene expression profiling has become a powerful research tool for understanding basic disease biology, and a powerful diagnostic tool for profiling cellular states during disease progression and in response to treatment. However, RNA purification , amplification, and labeling for microarrays, remain complex, time-consuming, labor-intensive, and expensive processes. The primary goal of this proposal is produce devices and systems that dramatically reduce this burden, so that it is no longer an imped iment to health-related research and diagnostics.