Amplifying mRNA Sequences from Small Amounts of Tissue

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DESCRIPTION (provided by applicant): Identifying differently expressed genes and their mRNA transcripts is a valuable starting point in understanding the molecular mechanisms underlying biological processes in normal and diseased states. Two widely used methods to facilitate these studies are Gene Expression Microarrays and Quantitative Reverse Transcriptase-linked Polymerase Chain Reaction. Although these are powerful techniques, they require microgram quantities of input RNA. In many cases, these amounts of RNA can be difficult or impossible to obtain. This is particularly the case when examining mRNA levels in primary cells, tissue biopsies, micro-dissected tissues and single cells. In these situations, a means to generate sufficient cDNA for multiple tests from small amounts of RNA for analysis would be very valuable. In the Phase I Research we propose to develop such a method, and utilize a set on non-random DNA primers for the synthesis and amplification of cDNA. This method will have advantages over current methods in terms of coverage of mRNA sequence representation, ability to work with partially degraded RNA, simplicity, time savings and cost. Proof of concept has been established and a commercialization plan is described.