Microfluidics-based in situ Padlock/Rolling Circle Amplification System for Counting Single DNA Molecules in a Cell
| العنوان: | Microfluidics-based in situ Padlock/Rolling Circle Amplification System for Counting Single DNA Molecules in a Cell |
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| المؤلفون: | Yuri Ishigaki, Kiichi Sato, Mats Nilsson, Arisa Kuroda, Kae Sato |
| المصدر: | Analytical Sciences. 30:1107-1112 |
| بيانات النشر: | Springer Science and Business Media LLC, 2014. |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | In situ, Cells, Microfluidics, Nanotechnology, DNA, Nucleic acid amplification technique, Analytical Chemistry, chemistry.chemical_compound, chemistry, Rolling circle replication, Humans, Molecule, Biological system, Nucleic Acid Amplification Techniques, HeLa Cells |
| الوصف: | 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. |
| تدمد: | 1348-2246 0910-6340 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::417e150a75c41fa8359e534d8eca2e4dTest https://doi.org/10.2116/analsci.30.1107Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....417e150a75c41fa8359e534d8eca2e4d |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 13482246 09106340 |
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