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

制备高质量的cDNA是进行基因功能分析的必要条件。然而，逆转录PCR是一种指数扩增方法，由于酶促反应的反应速率取决于底物的浓度，因此对大多数RNA分子产生偏倚。在体外转录和RT反应体系中加入假底物RNA(Chum-RNA)，建立了一种扩增超低量RNA的新方法。Chum-RNA增加了系统中底物的虚拟浓度，酶可以催化极少量RNA分子的反应。这项新技术使我们能够构建一个高质量的cDNA文库，该文库反映了单细胞RNA中mRNA分子的原始分布。在这里，我们报道该技术可以应用于微阵列分析和实时PCR方法。

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.