論文 - 詳細
| RRC ID | 68901 |
|---|---|
| 著者 | Kuroda A, Ishigaki Y, Nilsson M, Sato K, Sato K. |
| タイトル | Microfluidics-based in situ padlock/rolling circle amplification system for counting single DNA molecules in a cell. |
| ジャーナル | Anal Sci |
| Abstract |
In situ padlock/rolling circle amplification (RCA) is a method used to amplify, visualize, and quantify target DNA molecules in cells. However, the multiple reaction steps involved make this technique costly and cumbersome. We developed a novel, simplified, automated microfluidic system for RCA, and demonstrated its effectiveness by counting amplified mitochondrial DNA fragments in HeLa cells. After optimizing the volume of the reaction solutions and washing buffer composition, the product yield was equal to that obtained by the conventional manual method. The required volume of reagents was reduced to 10 μL, which is less than half the volume used in the conventional method. To the best of our knowledge, this is the first report of an automated microfluidic method for in situ padlock/RCA, which can be useful for making highly efficient pathological diagnoses. |
| 巻・号 | 30(12) |
| ページ | 1107-12 |
| 公開日 | 2014-1-1 |
| DOI | 10.2116/analsci.30.1107 |
| PII | DN/JST.JSTAGE/analsci/30.1107 |
| PMID | 25492458 |
| MeSH | Cells / metabolism* DNA / analysis* HeLa Cells Humans Microfluidics* Nucleic Acid Amplification Techniques* |
| IF | 2.049 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 1 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | HeLa(RCB0007) |