| 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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| 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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