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基于微芯片电泳的脱氧核糖核酸片段的浓缩和分离

徐中其*,廣川健   

  1. Department of Chemistry &Chemical Engineering, Graduate School of Engineering, Hiroshima University, Higashi-Hiroshima 739-8527, Japan
  • 收稿日期:2008-07-03 修回日期:2008-08-28 出版日期:2009-01-30 发布日期:1983-03-25
  • 通讯作者: 徐中其
  • 基金资助:

Preconcentration and separation of DNA fragments based on microchip electrophoresis

XU Zhongqi*, HIROKAWA Takeshi   

  1. Department of Chemistry &Chemical Engineering, Graduate School of Engineering, Hiroshima University, Higashi-Hiroshima 739-8527, Japan
  • Received:2008-07-03 Revised:2008-08-28 Online:2009-01-30 Published:1983-03-25
  • Contact: XU Zhongqi

PDF

583

被引次数

10

摘要： 采用超负荷电动供给(electrokinetic supercharging, EKS)预浓缩技术,在微芯片电泳(MCE)上对脱氧核糖核酸（DNA）片段进行浓缩和分离。EKS是集合样品电动进样(EKI)和过渡等速电泳(tITP)的一种在线浓缩方法。研究表明：采用该方法后,在40.5 mm长的单通道芯片上能够实现对低浓度样品的大量进样、浓缩和基线分离。在普通的紫外检测条件(检测波长为260 nm)下,对DNA片段的平均检出限(S/N=3)约为0.07 mg/L,仅为十字芯片上的微芯片电泳检出限的1/40。本文还对浓缩过程中的一些关键因素和定性分析进行了探讨。

关键词: DNA片段 , 超负荷电动供给, 电动进样, 过渡等速电泳, 微芯片电泳

Abstract: An online preconcentration method, electrokinetic supercharging (EKS) was used for the enrichment of DNA fragments based on microchip electrophoresis （MCE）. EKS is a process that combines electrokinetic injection (EKI) with transient isotachophoresis (tITP). The results demonstrated that the large volume of low concentration sample could be introduced, preconcentrated, and eventually separated on a single channel microchip with the whole length of 40.5 mm. The limit of detection (S/N=3) of DNA fragments was around 0.07 mg/L, effectively improved 40-fold by EKS preconcentration with the normal UV detection at 260 nm. Some important parameters for enhancing preconcentration and qualitative analysis were examined.

Key words: DNA fragments , electrokinetic injection, electrokinetic supercharging, transient isotachophoresis, microchip electrophoresis