Modeling and experimental characterization of peak tailing in DNA gel electrophoresis
Capillary electrophoresis (CE) is an efficient separation method in analytical chemistry. It exploits the difference in electrophoretic migration velocities between charged molecular species in aqueous or diluted polymer solution when an external electric field is applied to achieve separation. Desp...
| Main Authors: | , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
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2014
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| Online Access: | https://hdl.handle.net/10356/100135 http://hdl.handle.net/10220/24114 |
| _version_ | 1811693536988364800 |
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| author | Sun, Yi Kwok, Yien Chian Nguyen, Nam-Trung |
| author2 | School of Mechanical and Aerospace Engineering |
| author_facet | School of Mechanical and Aerospace Engineering Sun, Yi Kwok, Yien Chian Nguyen, Nam-Trung |
| author_sort | Sun, Yi |
| collection | NTU |
| description | Capillary electrophoresis (CE) is an efficient separation method in analytical chemistry. It exploits the difference in electrophoretic migration velocities between charged molecular species in aqueous or diluted polymer solution when an external electric field is applied to achieve separation. Despite the standard assumption that electrophoretic data obtained from pulse-loaded molecular species should have Gaussian peak shapes, experimentally observed peaks are frequently distorted or highly asymmetric. Interaction of charged species with the wall of the capillary is the primary source for serious band broadening and peak tailing. This paper reports a mathematical model for the peak profiles in capillary electrophoresis, taking adsorption on capillary wall into account. The model is based on the advection–diffusion equation, Langmuir second order kinetic equation and appropriate boundary conditions. It is applied to simulate the gel electrophoretic separation of the 11 fragment Φ X174-Hae III double stranded DNA ladders in a polymeric microchip. By using the migration velocities and diffusivities from the measurement, and properly selecting two fitting parameters, namely adsorption and desorption coefficients, the simulated peak shapes show remarkable similarity with the experimental electrophoretic results. The effect of adsorption and desorption coefficients are also investigated and the result shows that adsorption of analytes from the main analyte zone and desorption of these analytes appear to be the reasons of peak tailing, with the latter being the major cause. |
| first_indexed | 2024-10-01T06:53:15Z |
| format | Journal Article |
| id | ntu-10356/100135 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T06:53:15Z |
| publishDate | 2014 |
| record_format | dspace |
| spelling | ntu-10356/1001352020-03-07T13:19:28Z Modeling and experimental characterization of peak tailing in DNA gel electrophoresis Sun, Yi Kwok, Yien Chian Nguyen, Nam-Trung School of Mechanical and Aerospace Engineering National Institute of Education DRNTU::Science::Chemistry Capillary electrophoresis (CE) is an efficient separation method in analytical chemistry. It exploits the difference in electrophoretic migration velocities between charged molecular species in aqueous or diluted polymer solution when an external electric field is applied to achieve separation. Despite the standard assumption that electrophoretic data obtained from pulse-loaded molecular species should have Gaussian peak shapes, experimentally observed peaks are frequently distorted or highly asymmetric. Interaction of charged species with the wall of the capillary is the primary source for serious band broadening and peak tailing. This paper reports a mathematical model for the peak profiles in capillary electrophoresis, taking adsorption on capillary wall into account. The model is based on the advection–diffusion equation, Langmuir second order kinetic equation and appropriate boundary conditions. It is applied to simulate the gel electrophoretic separation of the 11 fragment Φ X174-Hae III double stranded DNA ladders in a polymeric microchip. By using the migration velocities and diffusivities from the measurement, and properly selecting two fitting parameters, namely adsorption and desorption coefficients, the simulated peak shapes show remarkable similarity with the experimental electrophoretic results. The effect of adsorption and desorption coefficients are also investigated and the result shows that adsorption of analytes from the main analyte zone and desorption of these analytes appear to be the reasons of peak tailing, with the latter being the major cause. 2014-10-23T08:21:58Z 2019-12-06T20:17:16Z 2014-10-23T08:21:58Z 2019-12-06T20:17:16Z 2006 2006 Journal Article Sun, Y., Kwok, Y. C., & Nguyen, N.-T. (2007). Modeling and experimental characterization of peak tailing in DNA gel electrophoresis. Microfluidics and nanofluidics, 3(3), 323-332. https://hdl.handle.net/10356/100135 http://hdl.handle.net/10220/24114 10.1007/s10404-006-0126-3 83251 en Microfluidics and nanofluidics © 2006 Springer-Verlag. 10 p. |
| spellingShingle | DRNTU::Science::Chemistry Sun, Yi Kwok, Yien Chian Nguyen, Nam-Trung Modeling and experimental characterization of peak tailing in DNA gel electrophoresis |
| title | Modeling and experimental characterization of peak tailing in DNA gel electrophoresis |
| title_full | Modeling and experimental characterization of peak tailing in DNA gel electrophoresis |
| title_fullStr | Modeling and experimental characterization of peak tailing in DNA gel electrophoresis |
| title_full_unstemmed | Modeling and experimental characterization of peak tailing in DNA gel electrophoresis |
| title_short | Modeling and experimental characterization of peak tailing in DNA gel electrophoresis |
| title_sort | modeling and experimental characterization of peak tailing in dna gel electrophoresis |
| topic | DRNTU::Science::Chemistry |
| url | https://hdl.handle.net/10356/100135 http://hdl.handle.net/10220/24114 |
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