| Project/Area Number |
21651085
|
|---|
| Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Applied genomics
|
|---|
| Research Institution | Osaka University |
|---|
Principal Investigator |
NOJIMA Hiroshi 大阪大学, 微生物病研究所, 教授 (30156195)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
OKUZAKI Daisuke 大阪大学, 微生物病研究所, 助教 (00346131)
|
|---|
| Project Period (FY) |
2009 – 2011
|
| Project Status |
Completed (Fiscal Year 2011)
|
| Budget Amount *help |
¥3,370,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥270,000)
Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2010: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2009: ¥1,300,000 (Direct Cost: ¥1,300,000)
|
|---|
| Keywords | PCR / Chum-RNA / 線形増幅 / mRNA増幅 / センス鎖 / DNAマイクロアレイ / 極微量 / リアルタイムPCR |
|---|
| 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.
|
