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Ferrofluid plug as valve and actuator for whole-cell PCR on chip

机译:铁磁塞作为阀和执行器,用于芯片上全细胞PCR

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摘要

Ferrofluid plug was first used as valve and actuator to circumvent the evaporation of aqueous fluid in LOC. Moreover, this dynamic plug catered for aqueous fluid expansion during its heating in the microchannel. Whole-cell polymerase chain reaction (PCR) protocol was successfully implemented in a magnetically actuated closed-loop PCR microchip system. Here, the ferrofluid plug was used to reduce the evaporation of the PCR sample in the thermal lysis step. A whole-cell PCR model was used to demonstrate this concept. The DNA plasmid, pUC19, was transformed into DH5a competent cells. Primer pairs were designed to amplify region of interest in pUC19. The transformants were cultured and selected. Whole-cell PCR was performed on intact bacteria to screen for the presence of pUC19, containing the region of interests. Serial dilutions demonstrated the limit of detection for this system to be four bacterial cells. PCR amplifications were verified with regular thermocyclers and results were comparable.
机译:铁磁流体塞首先用作阀门和执行器,以防止LOC中含水流体的蒸发。而且,该动态塞在其在微通道中加热期间迎合了水性流体膨胀。全细胞聚合酶链反应(PCR)协议已成功地在磁驱动的闭环PCR微芯片系统中实现。在此,铁磁塞用于减少热裂解步骤中PCR样品的蒸发。全细胞PCR模型用于证明这一概念。 DNA质粒pUC19被转化到DH5a感受态细胞中。设计引物对以扩增pUC19中的目的区域。培养并选择转化体。对完整细菌进行全细胞PCR,以筛选是否存在pUC19(包含目标区域)。连续稀释表明该系统的检测极限是四个细菌细胞。 PCR扩增已通过常规热循环仪验证,结果可比。

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