High throughput Analysis of Gene Expression in single cells utilizing Lab-disc
| Project/Area Number |
24510171
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Microdevices/Nanodevices
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| Research Institution | Soka University |
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Principal Investigator |
KUBO Izumi 創価大学, 工学部, 教授 (40214986)
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
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| Keywords | 単一細胞 / 発現遺伝子 / ラボディスク / Hot cell-direct RT-PCR / 発現遺伝子解析 / Jurkat Cell / Hot Cell-direct RT-PCR / 検出 |
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| Outline of Final Research Achievements |
Accurate measurement of fluorescent increase of each microchamber entrapping single cell was performed on the lab disc using fluorescent microscope before and after Hot cell-direct RT-PCR. In order to compare gene expression fluorescence increase of RT-PCR product of the gene was normalized by that of β-actin. It was optimized by setting the ratio of TAMRA probe for β-actin and FAM probe for the gene to 2 and concentration of primers were set to 0.2 μM.Fluorescent ration before to after PCR of GAPDH expression was largest at the concentration of FAM probe 0.4mM and TAMRA probe 0.8mM and the fluorescent ratio was 3.8, which was almost same as that of β-actin. As for CD95, IL-2 and p65, the largest fluorescent ratio was 3.4, 3.8 and 2.3 respectively. These results suggest that the gene expression difference in an individual cell can be evaluated by the proposed method.
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Report
(4 results)
Research Products
(32 results)
