Microfluidic Integration of Parallel Solid-Phase Liquid Chromatography
Microfluidic Integration of Parallel Solid-Phase Liquid Chromatography
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DOI：
10.1021/ac400163u
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发表时间：
2013-03-05
影响因子：
7.4
通讯作者：
Hansen, Carl L.
中科院分区：
文献类型：
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作者：
Huft, Jens;Haynes, Charles A.;Hansen, Carl L.
我们报告了基于最近开发的柱几何结构的完全集成的微流控色谱系统的开发，该系统允许在具有多层软光刻（MSL）集成阀的聚（二甲基硅氧烷）微流控装置中对高性能分离柱进行坚固的填料。并行高性能分离柱和片上管道的组合用于实现片上色谱的完全集成系统，包括自动样品上样、可编程梯度生成、分离、荧光检测和样品回收的所有步骤。我们利用强阴离子交换（AEX）树脂分离荧光标记的DNA和平行纯化逆转录聚合酶链反应（RT-PCR）扩增的小鼠免疫球蛋白基因可变区，证明了该系统。在非混相油流中平行回收样品具有样品稀释度低、回收率高的优点。在亚纳克DNA样品上进行核酸大小选择和恢复的能力为芯片上基因组学应用带来了希望，包括测序文库制备、克隆和诊断样品分离。
We report the development of a fully integrated microfluidic chromatography system based on a recently developed column geometry that allows for robust packing of high-performance separation columns in poly-(dimethylsiloxane) microfluidic devices having integrated valves made by multilayer soft lithography (MSL). The combination of parallel high-performance separation columns and on-chip plumbing was used to achieve a fully integrated system for on-chip chromatography, including all steps of automated sample loading, programmable gradient generation, separation, fluorescent detection, and sample recovery. We demonstrate this system in the separation of fluorescently labeled DNA and parallel purification of reverse transcription polymerase chain reaction (RT-PCR) amplified variable regions of mouse immunoglobulin genes using a strong anion exchange (AEX) resin. Parallel sample recovery in an immiscible oil stream offers the advantage of low sample dilution and high recovery rates. The ability to perform nucleic acid size selection and recovery on subnanogram samples of DNA holds promise for on-chip genomics applications including sequencing library preparation, cloning, and sample fractionation for diagnostics.