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结合电渗流与FTA纸用于微流控设备上的DNA分析

Combining Electro-Osmotic Flow and FTA Paper for DNA Analysis on Microfluidic Devices.

作者信息

Wimbles Ryan, Melling Louise M, Shaw Kirsty J

机构信息

Faculty of Science and Engineering, Manchester Metropolitan University, Chester Street, Manchester M1 5GD, UK.

出版信息

Micromachines (Basel). 2016 Jul 14;7(7):119. doi: 10.3390/mi7070119.

Abstract

FTA paper can be used to protect a variety of biological samples prior to analysis, facilitating ease-of-transport to laboratories or long-term archive storage. The use of FTA paper as a solid phase eradicates the need to elute the nucleic acids from the matrix prior to DNA amplification, enabling both DNA purification and polymerase chain reaction (PCR)-based DNA amplification to be performed in a single chamber on the microfluidic device. A disc of FTA paper, containing a biological sample, was placed within the microfluidic device on top of wax-encapsulated DNA amplification reagents. The disc containing the biological sample was then cleaned up using Tris-EDTA (TE) buffer, which was passed over the disc, via electro-osmotic flow, in order to remove any potential inhibitors of downstream processes. DNA amplification was successfully performed (from buccal cells, whole blood and semen) using a Peltier thermal cycling system, whereupon the stored PCR reagents were released during the initial denaturing step due to the wax barrier melting between the FTA disc and PCR reagents. Such a system offers advantages in terms of a simple sample introduction interface and the ability to process archived samples in an integrated microfluidic environment with minimal risk of contamination.

摘要

FTA 纸可用于在分析前保护各种生物样本,便于运输至实验室或进行长期存档存储。使用 FTA 纸作为固相可消除在 DNA 扩增前从基质中洗脱核酸的需求,使得 DNA 纯化和基于聚合酶链反应(PCR)的 DNA 扩增能够在微流控装置的单个腔室中进行。将一片含有生物样本的 FTA 纸放置在微流控装置内蜡包封的 DNA 扩增试剂上方。然后使用 Tris-EDTA(TE)缓冲液对含有生物样本的纸片进行清理,该缓冲液通过电渗流流过纸片,以去除下游过程的任何潜在抑制剂。使用珀耳帖热循环系统成功进行了 DNA 扩增(来自颊细胞、全血和精液),由于 FTA 纸片与 PCR 试剂之间的蜡屏障熔化,储存的 PCR 试剂在初始变性步骤中被释放。这样的系统在简单的样品引入界面以及在集成微流控环境中处理存档样本且污染风险最小方面具有优势。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72b7/6190317/2242a9281e5d/micromachines-07-00119-g001.jpg

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