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一种用于快速聚合酶链反应的圆形铁磁流体驱动微芯片。

A circular ferrofluid driven microchip for rapid polymerase chain reaction.

作者信息

Sun Y, Kwok Y C, Nguyen N T

机构信息

National Institute of Education, Nanyang Technological University, 1 Nanyang Walk, Singapore 637616.

出版信息

Lab Chip. 2007 Aug;7(8):1012-7. doi: 10.1039/b700575j. Epub 2007 Jun 13.

Abstract

In the past few years, much attention has been paid to the development of miniaturized polymerase chain reaction (PCR) devices. After a continuous flow (CF) PCR chip was introduced, several CFPCR systems employing various pumping mechanisms were reported. However, the use of pumps increases cost and imposes a high requirement on microchip bonding integrity due to the application of high pressure. Other significant limitations of CFPCR devices include the large footprint of the microchip and the fixed cycle number which is dictated by the channel layout. In this paper, we present a novel circular close-loop ferrofluid driven microchip for rapid PCR. A small ferrofluid plug, containing sub-domain magnetic particles in a liquid carrier, is driven by an external magnet along the circular microchannel, which in turn propels the PCR mixture through three temperature zones. Amplification of a 500 bp lambda DNA fragment has been demonstrated on the polymethyl methacrylate (PMMA) PCR microchip fabricated by CO(2) laser ablation and bonded by a low pressure, high temperature technique. Successful PCR was achieved in less than 4 min. Effects of cycle number and cycle time on PCR products were investigated. Using a magnet as the actuator eliminates the need for expensive pumps and provides advantages of low cost, small power consumption, low requirement on bonding strength and flexible number of PCR cycles. Furthermore, the microchip has a much simpler design and smaller footprint compared to the rectangular serpentine CFPCR devices. To demonstrate its application in forensics, a 16-loci short tandem repeat (STR) sample was successfully amplified using the PCR microchip.

摘要

在过去几年中,小型化聚合酶链反应(PCR)设备的发展备受关注。在连续流动(CF)PCR芯片问世后,有报道称出现了几种采用不同泵送机制的CFPCR系统。然而,泵的使用增加了成本,并且由于高压的应用,对微芯片键合完整性提出了很高的要求。CFPCR设备的其他显著局限性包括微芯片占地面积大以及由通道布局决定的固定循环次数。在本文中,我们展示了一种用于快速PCR的新型圆形闭环铁磁流体驱动微芯片。一个小的铁磁流体塞,在液体载体中包含亚域磁性颗粒,由外部磁铁沿着圆形微通道驱动,进而推动PCR混合物通过三个温度区。在通过CO₂激光烧蚀制造并用低压高温技术键合的聚甲基丙烯酸甲酯(PMMA)PCR微芯片上,已证明对500 bp的λDNA片段进行了扩增。在不到4分钟内实现了成功的PCR。研究了循环次数和循环时间对PCR产物的影响。使用磁铁作为驱动器无需昂贵的泵,并具有低成本、低功耗、对键合强度要求低以及PCR循环次数灵活的优点。此外,与矩形蛇形CFPCR设备相比,该微芯片设计更简单,占地面积更小。为了证明其在法医学中的应用,使用该PCR微芯片成功扩增了一个16位点短串联重复(STR)样本。

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