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Isotachophoresis of Nucleic Acids in Agarose Gel Rods


V. N. Kondratova, I. V. Botezatu, V. P. Shelepov, and A. V. Lichtenstein*

Blokhin Russian Cancer Research Center, Russian Academy of Medical Sciences, Kashirskoe Shosse 24, 115478 Moscow, Russia; fax: (495) 324-1205; E-mail: alicht@online.ru

* To whom correspondence should be addressed.

Received April 8, 2009; Revision received May 15, 2009
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

DOI: 10.1134/S0006297909110169