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Controlling electrophoretic trapping of circular DNA by addition of starch preparations to agarose gels

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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

  1. Bloemendal, H. (1960), J. Chromatogr. 3, 1–10.

    Article  CAS  Google Scholar 

  2. 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.

    Google Scholar 

  3. Buleon, A., Colonna, P., Planchot, V., and Ball, S. (1998), Int. J. Biol. Macromol. 23, 85–112.

    Article  CAS  Google Scholar 

  4. 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.

    Google Scholar 

  5. Imberty, A., Buleon, A., Tran, V., and Perez, S. (1991), Starch 43, 375–384.

    Article  CAS  Google Scholar 

  6. 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.

    Google Scholar 

  7. Serwer, P. and Allen, J. L. (1984), Biochemistry 23, 922–927.

    Article  CAS  Google Scholar 

  8. Mickel, S., Arena, V., and Bauer, W. (1977), Nucleic Acids Res. 4, 1465–1482.

    Article  CAS  Google Scholar 

  9. Levene, S. D. and Zimm, B. H. (1987), Proc. Natl. Acad. Sci. USA 84, 4054–4057.

    Article  CAS  Google Scholar 

  10. Serwer, P. and Hayes, S. J. (1987), Electrophoresis 8, 244–246.

    Article  CAS  Google Scholar 

  11. Åkerman, B. (1998), Biophys. J. 74, 3140–3151.

    Article  Google Scholar 

  12. Cole, K. D. and Åkerman, B. (2000), BioMacromolecules 1, 771–781.

    Article  CAS  Google Scholar 

  13. Arnott, S., Fulmer, A., Scott, W. E., Dea, I. C. M., Moorhouse, R., and Rees, D. A. (1974), J. Mol. Biol. 90, 269–284.

    Article  CAS  Google Scholar 

  14. 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.

    Google Scholar 

  15. Serwer, P. (1983), Electrophoresis 4, 375.

    Article  CAS  Google Scholar 

  16. Djabourov, M., Clark, A. H., Rowlands, D. W., and Ross-Murphy, S. B. (1989), Macromolecules 22, 180–188.

    Article  CAS  Google Scholar 

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Correspondence to Kenneth D. Cole.

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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

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