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使用小型化毛细管电泳系统结合开槽瓶阵列进样系统对DNA片段进行高通量分析。

High-throughput analysis of DNA fragments using a miniaturized CE system combined with a slotted-vial array sample introduction system.

作者信息

Fan Xiao-Feng, Li Qi, Wang Shi-Li, Xu Zhang-Run, Du Wen-Bin, Fang Qun, Fang Zhao-Lun

机构信息

Research Center for Analytical Sciences, Northeastern University, Shenyang, PR China.

出版信息

Electrophoresis. 2008 Dec;29(23):4733-8. doi: 10.1002/elps.200800287.

DOI:10.1002/elps.200800287
PMID:19053071
Abstract

An automated nanoliter sample introduction system was combined to a liquid-core waveguide (LCW)-based microfluidic CE system for high-throughput analysis of DNA fragments. The main component of the sample introduction system was a motor-driven plate, on which a circular array of bottom-slotted vials containing sample/buffer solutions was placed. A 7 cm-long LCW capillary served as both the sample probe and separation channel. The inlet terminal of the capillary could pass through the slots of the vials for electrokinetic sample introduction, and the capillary outlet was immersed in the solution of a reservoir, behind which a PMT facing directly to the outlet was positioned. A diode laser was used as excitation source for LCW LIF detection. Performance of the system was demonstrated through the separation of DNA fragments. Baseline separation was achieved for all 11 fragments of PhiX174-HaeIII digest DNA with a throughput of 33/h. Theoretical plate number for 603 bp fragment was 7.3x10(6)/m, corresponding to a plate height 0.14 microm. The detection limitation for 603 bp fragment was 0.4 ng/microL with a precision of 2.2% RSD for the peak height. Automated sample changing and introduction were achieved with only 0.3 nL gross sample consumption for each cycle.

摘要

将一个自动纳升进样系统与基于液芯波导(LCW)的微流控毛细管电泳系统相结合,用于DNA片段的高通量分析。进样系统的主要部件是一个电机驱动板,上面放置着一个圆形阵列的底部带槽小瓶,小瓶中装有样品/缓冲溶液。一根7厘米长的LCW毛细管既作为样品探针又作为分离通道。毛细管的入口端可以穿过小瓶的槽口进行电动进样,毛细管出口浸入一个储液器的溶液中,在储液器后面放置着一个直接面向出口的光电倍增管。使用二极管激光器作为LCW激光诱导荧光检测的激发源。通过对DNA片段的分离展示了该系统的性能。对PhiX174-HaeIII消化DNA的所有11个片段实现了基线分离,通量为每小时33次。603 bp片段的理论塔板数为7.3×10⁶/m,对应的塔板高度为0.14微米。603 bp片段的检测限为0.4 ng/μL,峰高的相对标准偏差(RSD)精度为2.2%。每个循环仅消耗0.3 nL总样品量就实现了自动进样和样品更换。

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