Performance of electrokinetic supercharging for high-sensitivity detection of DNA fragments in chip gel electrophoresis

被引:53
作者
Xu, ZQ
Nishine, T
Arai, A
Hirokawa, T
机构
[1] Hiroshima Univ, Grad Sch Engn, Dept Chem & Chem Engn, Higashihiroshima 7398527, Japan
[2] Shimadzu Co, Life Sci Lab, Anal & Measuring Instruments Div, Kyoto, Japan
关键词
chip gel electrophoresis; electrokinetic injection; electrokinetic supercharging; preconcentration; transient isotachophoresis;
D O I
10.1002/elps.200406061
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:3875 / 3881
页数:7
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