2011 Fiscal Year Final Research Report
Development and application of a novel technique that allows analysis on the gene expression of a single cell.
| Project/Area Number |
21651085
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied genomics
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| Research Institution | Osaka University |
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Principal Investigator |
NOJIMA Hiroshi 大阪大学, 微生物病研究所, 教授 (30156195)
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| Co-Investigator(Kenkyū-buntansha) |
OKUZAKI Daisuke 大阪大学, 微生物病研究所, 助教 (00346131)
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| Project Period (FY) |
2009 – 2011
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| Keywords | PCR / Chum-RNA / 線形増幅 / mRNA増幅 / センス鎖 |
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| Research Abstract |
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.
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