Capillary Electrophoresis of DNA in Hydroxyethylcellulose

doi:10.6023/A12110935

Acta Chimica Sinica ›› 2013, Vol. 71 ›› Issue (02): 265-270.DOI: 10.6023/A12110935 Previous Articles Next Articles

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羟乙基纤维素中毛细管电泳DNA分离特性研究

刘晨晨^a, 李振庆^a, 孟凡明^a, 倪一^a, 窦晓鸣^a, 山口佳则^b,a

a 教育部光学仪器与系统工程研究中心上海市现代光学系统重点实验室上海理工大学光电信息与计算机工程学院上海 200093;
b 大阪大学先进光子学中心大阪 565-0871

投稿日期:2012-11-19 发布日期:2012-12-25
通讯作者: 李振庆 E-mail:zhenqingli@163.com
基金资助:
国家自然科学基金(No. 21205078);高等学校博士学科点专项科研基金博导类资助课题(No. 20123120110002);上海理工大学教师创新能力建设(No.GDCX-Y-1205);上海市教委科研创新项目(No. 13YZ073);上海市科技发展基金(No. 10540500700)和上海市研究生创新基金(No. JWCXSL1202)资助项目. 本文部分受上海市重点学科项目第三期项目(S30502)资助.

Capillary Electrophoresis of DNA in Hydroxyethylcellulose

Liu Chenchen^a, Li Zhenqing^a, Meng Fanming^a, Ni Yi^a, Dou Xiaoming^a, Yamaguchi Yoshinori^b,a

a Engineering Research Center of Optical Instruments and System, Ministry of Education, Shanghai Key Laboratory of Modern Optical System, School of Optical-Electrical and Computer Engineering, University of Shanghai for Science and Technology, Shanghai 200093;
b Photonics Advanced Research Center, Osaka University, Osaka 565-0871

Received:2012-11-19 Published:2012-12-25
Supported by:
Project supported by the National Natural Science Foundation of China (No. 21205078), Research Fund for the Doctoral Program of Higher Education of China (No. 20123120110002), Teachers’ Innovation Ability Construction of University of Shanghai for Science and Technology (No. GDCX-Y-1205), the Innovation Program of Shanghai Municipal Education Commission (No. 13YZ073), Shanghai Committee of Science and Technology (No. 10540500700) and the Innovation Fund Project For Graduate Student of Shanghai (No. JWCXSL1202), and partly supported by the Leading Academic Discipline Project of Shanghai Municipal Government (S30502).

Using hydroxyethylcellulose (HEC) solution as polymer matrix, this work systematically studied the separation performance of 100 base pairs (bp) DNA ladder (100～1500 bp) by direct current electric field capillary electrophoresis (CE). In the present paper, we systematically investigated the influence of polymer concentration and molecular weight of HEC, electric field strength (E), the effective length (l_e) and the shape of the capillary, the temperature of the background electrolyte (BGE) on the separation performance of DNA. Furthermore, we compared the migration of DNA in polymer with the non-gel sieving model. Results show that: (1) When the concentration of HEC is above the entangled threshold c^*, the mobility difference increases with the growth of molecular weight, whereas the mobility of DNA decreases with the rise of concentration of HEC. (2) The resolution of adjacent DNA fragments linearly increases with the effective length (l_e) of the capillary when l_e ranges from 4 to 12 cm. (3) The mobility of DNA increases with the growth of area ratio R (S_lateral/S_section), and thus the separation performance improves. (4) The increase of BGE temperature strengthens the diffusion effect of DNA, thus increases the mobility, and deteriorates the resolution. Based on the results above, we separated the φ×174-Hirc II digest by CE in an optimal electrophoretic condition. Experiment shows that rapid separation of φ×174-Hirc II digest was realized with high resolution. In our experiment, the fused-silica capillary is coated by acrylamide, and the background electrolyte (BGE) used for the sieving matrix contains 0.5×Tris-borate-EDTA (TBE) and 1×SYBR Green I. The DNA sample was injected for 2.0 s at an E of 100 V/cm. The self-build CE device involved is reliable and showed some remarkable achievements previously. Such a study is beneficial to the realization of rapid and effective separation of DNA, and allows deep insight into DNA migration in the polymer matrix under constant electric field.

Key words: capillary electrophoresis, hydroxyethylcellulose, DNA

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Liu Chenchen, Li Zhenqing, Meng Fanming, Ni Yi, Dou Xiaoming, Yamaguchi Yoshinori. Capillary Electrophoresis of DNA in Hydroxyethylcellulose[J]. Acta Chimica Sinica, 2013, 71(02): 265-270.

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羟乙基纤维素中毛细管电泳DNA分离特性研究

Capillary Electrophoresis of DNA in Hydroxyethylcellulose

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