Biochemistry (Moscow)

, 74:1285 | Cite as

Isotachophoresis of nucleic acids in agarose gel rods

  • V. N. Kondratova
  • I. V. Botezatu
  • V. P. Shelepov
  • A. V. LichtensteinEmail author
Short Communications


A new method of electrophoresis (isotachophoresis in agarose gel rods) in which nucleic acid molecules are not separated but, oppositely, are brought together into one band, was elaborated. Heterogeneous in size DNA and RNA polymers present in a few milliliters of a solution at so low concentration that their isolation by other methods is hardly attainable and fraught with losses are brought together into one visible narrow band when put in a discontinuous electric field. Polynucleotides migrate in dilute (0.1%) semifluid agarose gel that permits easy quantitative isolation of the band of interest. Resulting DNA can be used directly in PCR. The suggested method for isolation of micro amounts of nucleic acids from dilute solutions can be applied to forensic and clinical research and cancer gene diagnostics by the analysis of fragmented circulating DNA from bodily fluids.

Key words

DNA isotachophoresis circulating DNA agarose gel DNA isolation PCR 



bromophenol blue



Supplementary material

10541_2009_9117_MOESM1_ESM.pdf (170 kb)
Supplementary material, approximately 170 KB.


  1. 1.
    Anker, P., Mulcahy, H., Chen, X. Q., and Stroun, M. (1999) Cancer Metastasis Rev., 18, 65–73.CrossRefPubMedGoogle Scholar
  2. 2.
    Nawroz, H., Koch, W., Anker, P., Stroun, M., and Sidransky, D. (1996) Nat. Med., 2, 1035–1037.CrossRefPubMedGoogle Scholar
  3. 3.
    Botezatu, I., Serdyuk, O., Potapova, G., Shelepov, V., Alechina, R., Molyaka, Y., Anan’ev, V., Bazin, I., Garin, A., Narimanov, M., Knysh, V., Melkonyan, H., Umansky, S., and Lichtenstein, A. (2000) Clin. Chem., 46, 1078–1084.PubMedGoogle Scholar
  4. 4.
    Sidransky, D. (2002) Nat. Rev. Cancer, 2, 210–219.CrossRefPubMedGoogle Scholar
  5. 5.
    Diehl, F., Schmidt, K., Choti, M. A., Romans, K., Goodman, S., Li, M., Thornton, K., Agrawal, N., Sokoll, L., Szabo, S. A., Kinzler, K. W., Vogelstein, B., and Diaz, L. A., Jr. (2008) Nat. Med., 14, 985–990.CrossRefPubMedGoogle Scholar
  6. 6.
    Garcia, V., Garcia, J., Silva, J., Pena, C., Dominguez, G., Lorenzo, Y., Diaz, R., Alonso, I., Colas, A., Hurtado, A., Sanchez, A., and Bonilla, F. (2008) J. Cancer Res. Clin. Oncol., 134, 1165–1171.CrossRefPubMedGoogle Scholar
  7. 7.
    Wong, B. C., and Lo, Y. M. (2003) Expert. Rev. Mol. Diagn., 3, 785–797.CrossRefPubMedGoogle Scholar
  8. 8.
    Shekhtman, E. M., Anne, K., Melkonyan, H. S., Robbins, D. J., Warsof, S. L., and Umansky, S. R. (2009) Clin. Chem., 55, 723–729.CrossRefPubMedGoogle Scholar
  9. 9.
    Lofton-Day, C., Model, F., Devos, T., Tetzner, R., Distler, J., Schuster, M., Song, X., Lesche, R., Liebenberg, V., Ebert, M., Molnar, B., Grutzmann, R., Pilarsky, C., and Sledziewski, A. (2008) Clin. Chem., 54, 414–423.CrossRefPubMedGoogle Scholar
  10. 10.
    Melkonyan, H. S., Feaver, W. J., Meyer, E., Scheinker, V., Shekhtman, E. M., Xin, Z., and Umansky, S. R. (2008) Ann. N. Y. Acad. Sci., 1137, 73–81.CrossRefPubMedGoogle Scholar
  11. 11.
    Kagan, J., Srivastava, S., Barker, P. E., Belinsky, S. A., and Cairns, P. (2007) Cancer Res., 67, 4545–4549.CrossRefPubMedGoogle Scholar
  12. 12.
    Rago, C., Huso, D. L., Diehl, F., Karim, B., Liu, G., Papadopoulos, N., Samuels, Y., Velculescu, V. E., Vogelstein, B., Kinzler, K. W., and Diaz, L. A., Jr. (2007) Cancer Res., 67, 9364–9370.CrossRefPubMedGoogle Scholar
  13. 13.
    Hunter, M. P., Ismail, N., Zhang, X., Aguda, B. D., Lee, E. J., Yu, L., Xiao, T., Schafer, J., Lee, M. L., Schmittgen, T. D., Nana-Sinkam, S. P., Jarjoura, D., and Marsh, C. B. (2008) PLoS ONE, 3, e3694.CrossRefPubMedGoogle Scholar
  14. 14.
    Mitchell, P. S., Parkin, R. K., Kroh, E. M., Fritz, B. R., Wyman, S. K., Pogosova-Agadjanyan, E. L., Peterson, A., Noteboom, J., O’Briant, K. C., Allen, A., Lin, D. W., Urban, N., Drescher, C. W., Knudsen, B. S., Stirewalt, D. L., Gentleman, R., Vessella, R. L., Nelson, P. S., Martin, D. B., and Tewari, M. (2008) PNAS, 105, 10513–10518.CrossRefPubMedGoogle Scholar
  15. 15.
    Beck, J., Urnovitz, H. B., Riggert, J., Clerici, M., and Schutz, E. (2009) Clin. Chem., 55, 730–738.CrossRefPubMedGoogle Scholar
  16. 16.
    Schmidt, B., Weickmann, S., Witt, C., and Fleischhacker, M. (2005) Clin. Chem., 51, 1561–1563.CrossRefPubMedGoogle Scholar
  17. 17.
    Herrera, L. J., Raja, S., Gooding, W. E., El Hefnawy, T., Kelly, L., Luketich, J. D., and Godfrey, T. E. (2005) Clin. Chem., 51, 113–118.CrossRefPubMedGoogle Scholar
  18. 18.
    Sozzi, G., Conte, D., Leon, M., Ciricione, R., Roz, L., Ratcliffe, C., Roz, E., Cirenei, N., Bellomi, M., Pelosi, G., Pierotti, M. A., and Pastorino, U. (2003) J. Clin. Oncol., 21, 3902–3908.CrossRefPubMedGoogle Scholar
  19. 19.
    Abelev, G. I. (1979) Meth. Cancer Res., 18, 2–38.Google Scholar
  20. 20.
    Abelev, G. I., and Karamova, E. R. (1996) Biochemistry (Moscow), 61, 1413–1421.Google Scholar
  21. 21.
    Abelev, G. I., and Karamova E. R. (1982) Zh. Vsesoyuz. Khim. O-va im. D. I. Mendeleeva, 27, 429–475.Google Scholar
  22. 22.
    Kondratova, V. N., Serd’uk, O. I., Shelepov, V. P., and Lichtenstein, A. V. (2005) Biotechniques, 39, 695–699.CrossRefPubMedGoogle Scholar
  23. 23.
    Ma, H., and Difazio, S. (2008) Biotechniques, 44, 921–923.CrossRefPubMedGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • V. N. Kondratova
    • 1
  • I. V. Botezatu
    • 1
  • V. P. Shelepov
    • 1
  • A. V. Lichtenstein
    • 1
    Email author
  1. 1.Blokhin Russian Cancer Research CenterRussian Academy of Medical SciencesMoscowRussia

Personalised recommendations