Integrated microfluidic chip for rapid DNA digestion and time-resolved capillary electrophoresis analysis.
Integrated microfluidic chip for rapid DNA digestion and time-resolved capillary electrophoresis analysis.
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DOI：
10.1063/1.3654950
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发表时间：
2012-03
期刊：
影响因子：
3.2
通讯作者：
Che-Hsin Lin;Y. Wang;Lung-Ming Fu
中科院分区：
文献类型：
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作者：
Che-Hsin Lin;Y. Wang;Lung-Ming Fu
提出了一种用于DNA快速消化和时间分辨毛细管电泳法分析的集成微流控芯片。该芯片包括两个用于DNA浓缩和纯化的凝胶填充小室，一个用于DNA/限制性内切酶混合的T型微混合器，一个用于DNA消化反应的蛇形通道，以及一个用于在线毛细管电泳法分析的CE通道。DNA和限制性内切酶使用夹持开关直流场进行电镜学混合。实验和数值结果表明，当驱动电压为90V/cm，开关频率为4 Hz时，在距T结1 mm的范围内，混频性能可达97%。连续混合消化和毛细管电泳法操作清楚地呈现了不同CE运行中对φx-174DNA消化的变化。时间分辨电泳图显示，所提出的装置能够在24分钟内浓缩和分析包含11个片段的φx-174DNA样本。总体而言，这项研究的结果表明，所提出的微流控芯片为DNA消化和CE分析应用提供了一种快速有效的工具。
An integrated microfluidic chip is proposed for rapid DNA digestion and time-resolved capillary electrophoresis (CE) analysis. The chip comprises two gel-filled chambers for DNA enrichment and purification, respectively, a T-form micromixer for DNA/restriction enzyme mixing, a serpentine channel for DNA digestion reaction, and a CE channel for on-line capillary electrophoresis analysis. The DNA and restriction enzyme are mixed electroomostically using a pinched-switching DC field. The experimental and numerical results show that a mixing performance of 97% is achieved within a distance of 1 mm from the T-junction when a driving voltage of 90 V/cm and a switching frequency of 4 Hz are applied. Successive mixing digestion and capillary electrophoresis operation clearly present the changes on digesting φx-174 DNA in different CE runs. The time-resolved electropherograms show that the proposed device enables a φx-174 DNA sample comprising 11 fragments to be concentrated and analyzed within 24 min. Overall, the results presented in this study show that the proposed microfluidic chip provides a rapid and effective tool for DNA digestion and CE analysis applications.