Development and application of a novel technique that allows analysis on the gene expression of a single cell.

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Preparation of high-quality cDNA is needed for functional analysis of gene function. However, reverse transcription PCR, an exponential amplification method causes a bias to majority of RNA molecules, because the reaction rate of enzymatic reaction depends on the concentration of the substrates. We developed a novel method for amplification of ultralow amount RNA by addition of dummy substrate RNA(Chum-RNA) into in vitro transcription and RT reaction system. Chum-RNA increases virtual concentration of substrate in the system, and the enzymes can catalyze the reaction for a very small number of RNA molecules. This novel technology enabled us to construct a high-quality cDNA library which reflects the original distribution of mRNA molecules from a single cell amount of RNA. Here, we report that this technology can be applied to Microarray analysis and Real time PCR method.