Fully integrated PCR-capillary electrophoresis microsystem for DNA analysis
Fully integrated PCR-capillary electrophoresis microsystem for DNA analysis
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DOI：
10.1039/b109031n
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发表时间：
2001-01-01
期刊：
影响因子：
6.1
通讯作者：
Mathies, RA
中科院分区：
文献类型：
--
作者：
Lagally, ET;Emrich, CA;Mathies, RA
已经构建了一个完全集成的基因组分析微系统，包括微制造加热器，温度传感器和直接连接到毛细管电泳分离通道的PCR室。阀门和疏水通风口提供控制和无传感器的样品定位和固定到200 nL PCR室。使用微制造加热和温度传感元件将PCR反应的加热和冷却速率提高到20℃（-1）。扩增的PCR产物，用插层荧光染料在柱上标记，注射到凝胶填充的毛细管中进行电泳分析。成功的性别决定使用多重PCR反应从人类基因组DNA证明在不到15分钟。该装置是向用于法医和即时分子医学诊断的微制造基因组微处理器迈出的重要一步。
A fully integrated genomic analysis microsystem including microfabricated heaters, temperature sensors, and PCR chambers directly connected to capillary electrophoretic separation channels has been constructed. Valves and hydrophobic vents provide controlled and sensorless sample positioning and immobilization into 200 nL PCR chambers. The use of microfabricated heating and temperature sensing elements improves the heating and cooling rates for the PCR reaction to 20 degreesC s(-1). The amplified PCR product, labeled on-column with an intercalating fluorescent dye, is injected into the gel-filled capillary for electrophoretic analysis. Successful sex determination using a multiplex PCR reaction from human genomic DNA is demonstrated in less than 15 min. This device is an important step toward a microfabricated genomic microprocessor for use in forensics and point-of-care molecular medical diagnostics.