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Lab-on-capillary: a rapid, simple and quantitative genetic analysis platform integrating nucleic acid extraction, amplification and detection

机译:毛细管 - 毛细管:快速,简单,定量的遗传分析平台,整合核酸提取,扩增和检测

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摘要

In this work, we describe for the first time a genetic diagnosis platform employing a poly-diallyldimethylammonium chloride (PDDA)-modified capillary and a liquid-based thermalization system for rapid, simple and quantitative DNA analysis with minimal user interaction. Positively charged PDDA is modified on the inner surface of the silicon dioxide capillary by using an electrostatic self-assembly approach that allows the negatively charged DNA to be separated from the lysate in less than 20 seconds. The capillary loaded with the PCR mix is incorporated in the thermalization system, which can achieve on-site real-time PCR. This system is based on the circulation of pre-heated liquids in the chamber, allowing for highspeed thermalization of the capillary and fast amplification. Multiple targets can be simultaneously analysed with multiplex spatial melting. Starting with live Escherichia coli (E. coli) cells in milk, as a realistic sample, the current method can achieve DNA extraction, amplification, and detection within 40 min.
机译:在这项工作中,我们首次描述了采用聚二烯丙基甲基氯化铵(PDDA)制定的毛细血管(PDDA)的遗传诊断平台,以及用于快速,简单,定量的DNA分析的基于液体的热化系统,具有最小的用户相互作用。通过使用静电自组装方法在二氧化硅毛细管的内表面上改变带正电的PDDA,该静电自组装方法允许带负电荷的DNA在小于20秒的裂解物中与裂解物分离。加载有PCR混合物的毛细管掺入热化系统中,可以实现现场实时PCR。该系统基于腔室中预热液体的循环,允许高速热化毛细管和快速放大。通过多路复用空间熔化可以同时分析多个靶标。从牛奶中的活大肠杆菌(大肠杆菌)细胞开始,作为现实样品,目前的方法可以在40分钟内实现DNA提取,扩增和检测。

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