Simultaneous extraction and analyses of RNA and DNA from single cells
Project/Area Number |
26630052
|
Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
Allocation Type | Multi-year Fund |
Research Field |
Fluid engineering
|
Research Institution | Kyoto University |
Principal Investigator |
|
Co-Investigator(Kenkyū-buntansha) |
小寺 秀俊 京都大学, 工学研究科, 教授 (20252471)
|
Research Collaborator |
Santiago Juan G. Stanford University, 教授
NISHIKII Hidekazu Stanford University, Visiting Scholar
|
Project Period (FY) |
2014-04-01 – 2017-03-31
|
Project Status |
Completed (Fiscal Year 2016)
|
Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
|
Keywords | マイクロ流路 / 電気泳動 / 一細胞 / RNA / DNA / シークエンシング / 電気穿孔 / 前処理 / マイクロ流体 / 動電現象 / 等速電気泳動法 / タンパク / PCR |
Outline of Final Research Achievements |
In this project, we have developed a novel extraction technique of RNA and DNA from single cells with electrical lysis and electrokinetics and demonstrated it with integrated analyses on RNA and DNA. The technique leverages a microfluidic system to enable selective lysis of cytoplasmic membrane in less than 200 ms and completes extraction and separation between cytoplasmic RNA and a nucleus in less than one minute. The extracted RNA and DNA molecules showed high quality to perform high-throughput sequencing analyses. We also demonstrated our techniques with integrated nuclear and cytoplasmic RNA sequencing analyses and proposed a new strategy to study RNA transport at single cell resolution.
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Report
(4 results)
Research Products
(22 results)