Xu Zhongqi, Nishine Tsutomu, Arai Akihiro, Hirokawa Takeshi
Applied Chemistry, Department of Chemistry and Chemical Engineering, Graduate School of Engineering, Hiroshima University, Higashi-hiroshima 739-8527, Japan.
Electrophoresis. 2004 Nov;25(21-22):3875-81. doi: 10.1002/elps.200406061.
Chip gel electrophoresis was explored for high-sensitivity detection of DNA by combining electrokinetic injection with transient isotachophoresis preconcentration (here named electrokinetic supercharging (EKS)). Low concentrations (0.2 mg/L) of DNA sample could be detected without fluorescence labeling using a conventional UV detector (at 260 nm). On a single-channel microchip, identification of PCR product was performed by exploiting both external and internal calibration methods. The deviation between the two calibration methods was about 3.6%, and the identified DNA fragment size matched with the predicted size of the template DNA. On the cross microchip the EKS preconcentration has also been achieved when changing the injection reservoir differing from the one used previously. The procedure was computer-simulated and the influence of the voltage applied to two-side reservoirs on sample preconcentration and dilution was also discussed.
通过将电动进样与瞬态等速电泳预富集（此处称为电动增压（EKS））相结合，探索了芯片凝胶电泳用于DNA的高灵敏度检测。使用传统的紫外检测器（260nm），无需荧光标记即可检测低浓度（0.2mg/L）的DNA样品。在单通道微芯片上，利用外部和内部校准方法对PCR产物进行鉴定。两种校准方法之间的偏差约为3.6%，鉴定出的DNA片段大小与模板DNA的预测大小相符。在交叉微芯片上，当改变进样储液器与先前使用的不同时，也实现了EKS预富集。对该过程进行了计算机模拟，并讨论了施加到两侧储液器上的电压对样品预富集和稀释的影响。
