Application of Lab-Disk to high-throughput genomic profiling of single cells
| Project/Area Number |
21510127
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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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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| Co-Investigator(Renkei-kenkyūsha) |
NAGAI Hidenori 産業技術総合研究所, 健康工学研究センター, 研究員 (40357596)
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2011: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2009: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | マイクロセンサー / ハイスループット / ラボディスク / Jurkat Cell / GAPDH / Hot Cell Direct RT-PCR / ナノリッターPCR / 蛍光プローブ / 単一細胞分離 / マイクロチャンバー / Hot Cell direct RT-PCR / ディスク / Double dye probe / 単一細胞 / RT-PCR / m-RNA / ナノリッター |
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| Research Abstract |
We have developed a CD-shaped device(Lab-disk) to isolate single cell from a large number of cells simply and rapidly. In order to perform PCR or RT-PCR on the device, the device was fabricated from silicon and glass not to evaporate the entrapped solution in the microchambers in this study. Amplification of Salmonella specific invA gene form single cell was confirmed on the disk at 200 cells/μl of Salmonella enterica after single cell isolation and PCR. To realize q-PCR of Jurkat cell, hot cell-direct RT-PCR was enabled by using Tth DNA Polymerase, which is a thermo-stable polymerase and has high reverse transcription activity. After cell isolation and successive hot cell-direct RT-PCR on the device, fluorescent signals from RT-PCR products for single cells were detected at 200 cells/μl of Jurkat cell or less.
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Report
(4 results)
Research Products
(70 results)
