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

被引:5
|
作者
Wimbles, Ryan [1 ]
Melling, Louise M. [1 ]
Shaw, Kirsty J. [1 ]
机构
[1] Manchester Metropolitan Univ, Fac Sci & Engn, Chester St, Manchester M1 5GD, Lancs, England
来源
MICROMACHINES | 2016年 / 7卷 / 07期
关键词
microfluidic; DNA purification; DNA amplification; FTA (R) paper; electro-osmotic flow; EXTRACTION; SYSTEM; IDENTIFICATION; PURIFICATION; BLOOD; PCR;
D O I
10.3390/mi7070119
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
FTA (R) 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 (R) 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 (R) 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 (R) 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.
引用
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页数:9
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