On-line pre-concentration and UV determination of DNA fragments by dynamic coating capillary electrophoresis and its application to detection of genetically modified oilseed rape based on PCR

文献类型: 外文期刊

第一作者: Chen, Hong

作者: Chen, Hong;Wu, Yu-Hua;Song, Dan-Yang;Zhang, Wen;Dong, Xu-Yan;Li, Pei-Wu;Lu, Chang-Ming

作者机构:

关键词: capillary electrophoresis;dynamic coating;DNA fragment;UV detection;transgenic rapeseed;genetically modified organisms;LASER-INDUCED FLUORESCENCE;POLYMERASE-CHAIN-REACTION;GEL-ELECTROPHORESIS;RESTRICTION FRAGMENTS;REACTION-PRODUCTS;ISOTACHOPHORETIC PRECONCENTRATION;MODIFIED MAIZE;FOOD ANALYSIS;SEPARATION;POLYACRYLAMIDE

期刊名称:MICROCHEMICAL JOURNAL ( 影响因子:4.821; 五年影响因子:4.364 )

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收录情况: SCI

摘要: In this work, it was demonstrated that on-line pre-concentration and separation of DNA fragments within bared silica column by dynamic coating capillary electrophoresis and UV detection. The DNA fragments were pre-concentrated with long electrokinetic injecting time (99 s), peak height increased dramatically as a function of injection time, especially for shorter length DNA. The concentration sensitivity of DNA fragments can be improved from 20- to 100-fold relative to a normal injection (5 s). The electro-osmotic flow (EOF) and DNA-wall interactions within the capillary were eliminated effectively by dynamic coating method. Employing 0.5% poly(ethylene oxide) (PEO) in Tris-phosphate-EDTA (TBE) buffer as sieving matrix, DNA fragments, ranging from 11 to 657 bp, were separated within 20 min. The linear coefficient of linear relation between the migration and DNA length is 0.999. The DNA fragments amplified from transgenic oilseed rape by polymerase chain reaction (PCR) were separated and detected by this method, demonstrating the potential use of this method for effective DNA analysis and detection of genetically modified organisms (GMO). (C) 2006 Elsevier B.V. All rights reserved.

分类号: O655

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