Study on the Mobilities with Replaceable Linear Polyacrylamide (LPA) Gel Electrophoresis

Abstract:

The replaceable Linear polyacrylamide(LPA) gel was used as the sieving medium; An extensive series of experiments has been performed using DNA fragments ranging in size from 80 to 587 base pairs with laser induced fluorescence detection(LIFD). The electric field was changed from 100V/cm to 375V/cm. The mobilities with LPA in different electric field strength were studied. Although the LPA sieving matrix is commonly used in biological chemistry, these mobilities have not previously been reported. It was different from with the cross-link polyacrylamide and agar gels. The new modification of Ogston model was initiated. This work complements our systematic study on electrophoretic mobilities under different conditions, and will help to further optimize and improve high performance DNA separation in microfluidic applications.

• Periodical: Advanced Materials Research (Volume 681)
• Pages: 16-20
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.681.16
• Online since: April 2013
• Authors: Jie Wang, Li Na Guo, Li Qiang Wang, Yan Shi, Hua Zheng, Zu Kang Lu
• Keywords: Electric Field, Linear Polyacrylamide (LPA), Mobility

References

[1] K. E. Allan, C. E. Lenehan, D. A. Khodakov, et. al. High-performance capillary electrophoretic separation of double-stranded oligonucleotides using a poly- (ethylpyrrolidine methacrylate-co-methyl methacrylate)-coated capillary, ELECTROPHORESIS, 33(2012).

DOI: 10.1002/elps.201100514

[2] E. A. S. Doherty, C. W. Kan, B. M. Paegel, et. al. Sparsely Cross-Linked "Nanogel" Matrixes as Fluid, Mechanically Stabilized Polymer Networks for High-Throughput Microchannel DNA Sequencing, Anal. Chem., 76(2004)5249-5256.

DOI: 10.1021/ac049721x

[3] E. H. Turner, K. Lauterbach, H. R. Pugsley, et. al. Detection of green fluorescent protein in a single bacterium by capillary electrophoresis with laser-induced fluorescence, Anal Chem, 79(2007)778-781.

DOI: 10.1021/ac061778r

[4] A. Kotani, M. A. Witek, J. K. Osiri, H. Wang, et. al. EndoV/DNA ligase mutation scanning assay using microchip capillary electrophoresis and dual-color laser-induced fluorescence detection, Analytical Methods, 4(2012)58-64.

DOI: 10.1039/c1ay05366c

[5] G. W. Slater and H. L. Guo, An exactly solvable Ogston model of gel electrophoresis: I. The role of the symmetry and randomness of the gel structure, Electrophoresis, 17(1996) 977-988.

DOI: 10.1002/elps.1150170604

[6] Z. Shen, X. Liu, X. Zhou, et. al. Quantitative evaluation of the interaction between netropsin and double stranded oligodeoxynucleotides by microfabricated capillary array electrophoresis, J Sep Sci, 30(2007)1544-1548.

DOI: 10.1002/jssc.200600530

[7] M. C. Ruiz-Martinez, J. Berka, A. Belenkii, et. al. DNA sequencing by capillary electrophoresis with replaceable linear polyacrylamide and laser-induced fluorescence detection, Anal Chem, 65(1993)2851-2858.

DOI: 10.1021/ac00068a023

[8] T. Kaneta, T. Ogura, S. Yamato, and T. Imasaka, Band broadening of DNA fragments isolated by polyacrylamide gel electrophoresis in capillary electrophoresis, Journal of Separation Science, 35(2012) 431-435.

DOI: 10.1002/jssc.201100909

[9] P. D. B. H. Z. Oscar J. Lumpkin, Theory of gel electrophoresis of DNA, Biopolymers, 24(1985)1573-1593.

[10] J. R. J. N. C. T. M. L. Gary W. Slater, Quantitative analysis of the three regimes of DNA electrophoresis in agarose gels, Biopolymers, 27(1988)509-524.

DOI: 10.1002/bip.360270311

[11] J. N. Gary W. Slater, The biased reptation model of DNA gel electrophoresis: Mobility vs molecular size and gel concentration, Biopolymers, 28(1989)1781-1791.

DOI: 10.1002/bip.360281011

[12] J. A. Luckey and L. M. Smith, A model for the mobility of single-stranded DNA in capillary gel electrophoresis, Electrophoresis, 14(1993) 492-501.

DOI: 10.1002/elps.1150140176