Abstract
Starch preparations were added to agarose gels to enhance the electrophoretic trapping of circular plasmid DNA. The critical voltages required to trap the open circular (OC) and the supercoiled (SC) forms of a 13.1-kbp plasmid were measured in gels composed of agarose and added starch preparations. Modified starch preparations reduced the critical voltage required to trap the OC form of the plasmid to approximately one-third of the control value (in 1% agarose gels). Amylose (a fraction of starch with a low amount of branching) also reduced the critical voltage to trap the OC form in a similar manner. The critical voltage to trap the SC form of the plasmid was not significantly reduced by the starch preparations. The capacity to trap OC DNA was increased by the addition of higher amounts of the starch preparations added to the gels. Field inversion gel electrophoresis was used to characterize the length of the traps in the gels. The starch preparations and amylose increased the trap lengths approximately twofold. The increased trap length correlated with the decreased critical voltage required to trap the OC form of the 13.1-kbp plasmid.
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References
Bloemendal, H. (1960), J. Chromatogr. 3, 1–10.
Gordon, A. H. (1969), in Laboratory Techniques in Biochemistry and Molecular Biology, vol. 1, Work, T. S. and Work, E., eds., North-Holland Publishing, Amsterdam, pp. 1–150.
Buleon, A., Colonna, P., Planchot, V., and Ball, S. (1998), Int. J. Biol. Macromol. 23, 85–112.
Whistler, R. L. and Daniel, J. R. (1984), in Starch: Chemistry and Function, 2nd ed., Whistler, R. L., BeMiller, J. N., and Paschall, E. F., eds., Academic Press, Orlando, pp. 153–182.
Imberty, A., Buleon, A., Tran, V., and Perez, S. (1991), Starch 43, 375–384.
Stellwagen, N. C. (1987), in Adv. in Electrophoresis, vol. 1, Chrambach, A., Dunn, M. J., and Radola, B. J., eds., VCH, Weinheim, pp. 177–228.
Serwer, P. and Allen, J. L. (1984), Biochemistry 23, 922–927.
Mickel, S., Arena, V., and Bauer, W. (1977), Nucleic Acids Res. 4, 1465–1482.
Levene, S. D. and Zimm, B. H. (1987), Proc. Natl. Acad. Sci. USA 84, 4054–4057.
Serwer, P. and Hayes, S. J. (1987), Electrophoresis 8, 244–246.
Åkerman, B. (1998), Biophys. J. 74, 3140–3151.
Cole, K. D. and Åkerman, B. (2000), BioMacromolecules 1, 771–781.
Arnott, S., Fulmer, A., Scott, W. E., Dea, I. C. M., Moorhouse, R., and Rees, D. A. (1974), J. Mol. Biol. 90, 269–284.
Kirkpatrick, F. H. (1990), in Electrophoresis of Large DNA Molecules: Theory and Applications, vol. 9, Lai, E. and Birren, B. W., eds., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 9–22.
Serwer, P. (1983), Electrophoresis 4, 375.
Djabourov, M., Clark, A. H., Rowlands, D. W., and Ross-Murphy, S. B. (1989), Macromolecules 22, 180–188.
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Certain commercial equipment, instruments, or materials are identified in this article to specify adequately the experimental procedures. Such identification does not imply recommendation by National Institute of Standards and Technology, nor does it imply that the materials or equipment are necessarily the best available for the purpose.
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Cole, K.D., Tellez, C.M. & Nguyen, R. Controlling electrophoretic trapping of circular DNA by addition of starch preparations to agarose gels. Appl Biochem Biotechnol 95, 31–43 (2001). https://doi.org/10.1385/ABAB:95:1:31
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DOI: https://doi.org/10.1385/ABAB:95:1:31