Separation of long DNA fragments by inversion field capillary electrophoresis |
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| Authors: | Zhenqing Li Xiaoming Dou Yi Ni Yoshinori Yamaguchi |
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| Affiliation: | (1) Institute of Optical-Electrical Engineering, University of Shanghai for Science and Technology, 516 Jungong Rd., Yangpu, Shanghai, 200093, China;(2) Department of Physics, Optical engineering, Shanghai Jiao Tong University, 800 Dongchuan Rd., Minhang, Shanghai, 200240, China;(3) Institute for Biomedical Engineering, Consolidated Research Institute for Advanced Science and Medical Care (ASMeW), Waseda University, 513 Wasedatsurumaki-cho, Shinjuku-ku Tokyo, 162-0041, Japan;(4) Photonics Advanced Research Center (PARC), Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita Osaka, 565-0871, Japan; |
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| Abstract: | This study reports improved pulsed field capillary electrophoresis (PFCE) for separation of large DNA ladders. Important analytical conditions, including gel polymer concentration, ratio of forward to backward pulse duration, and separation potential, were investigated for their effects on the separation performance of DNA ranging in size from 0.1 to 10.0 kilo base pairs (kbp). Results show that DNA fragments from 0.1 to 8.0 kbp can be resolved with high resolution, simultaneously, in a short time. The ratio of forward to backward pulse duration affects the separation performance for DNA fragments greater than 1.5 kbp, and 3 or 4 is the optimum value of the ratio for separation of DNA up to 10 kbp. Furthermore, the separations that were obtained with 74–19,329 bp λ-DNA restriction fragments clearly demonstrate a dramatic improvement in the separation time and resolution over the conventionally used square-wave PFCE. The inversion field capillary electrophoresis reported here may help enable future DNA analysis studies to be performed quickly and effectively. |
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