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Microbead-based rolling circle amplification in a microchip for sensitive DNA detection
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Microbead-based rolling circle amplification in a microchip for sensitive DNA detection

Kae Sato, Atsuki Tachihara, Björn Renberg, Kazuma Mawatari, Kiichi Sato, Yuki Tanaka, Jonas Jarvius, Mats NilssonTakehiko Kitamori
Lab on a Chip, 卷.10(10), 頁碼.1262-1266
2010

摘要

Bioengineering Biochemistry Chemistry (all) Biomedical Engineering
The sensitive detection and quantification of DNA targets in the food industry and in environmental and clinical settings are issues of utmost importance in ensuring contamination-free food, monitoring the environment, and battling disease. Selective probes coupled with powerful amplification techniques are therefore of major interest. In this study, we set out to create an integrated microchemical chip that benefits from microfluidic chip technology in terms of sensitivity and a strong detection methodology provided jointly by padlock probes and rolling circle amplification (RCA). Here, we have integrated padlock probes and RCA into a microchip. The chip uses solid phase capture in a microchannel to enable washing cycles and decrease analytical area, and employs on-bead RCA for single-molecule amplification and detection. We investigated the effects of reagent concentration and amount of padlock probes, and demonstrated the feasibility of detecting Salmonella. © 2010 The Royal Society of Chemistry.

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