Separation of 1–23-kb complementary DNA strands by urea–agarose gel electrophoresis |
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| Authors: | éva Hegedüs Endre Kókai Alexander Kotlyar Viktor Dombrádi Gábor Szabó |
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| Affiliation: | 1.Department of Biophysics and Cell Biology, 2.Department of Medical Chemistry, University of Debrecen, 4012 Debrecen, Nagyerdei krt. 98, Hungary, 3.Department of Biochemistry, George S. Wise Faculty of Life Sciences and Nanotechnology Center, Tel Aviv University, Ramat Aviv 69978, Israel and 4.HAS Cell Biology and Signalling Research Group, Medical and Health Science Center, University of Debrecen, 4012 Debrecen, Nagyerdei krt. 98, Hungary |
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| Abstract: | Double-stranded (ds), as well as denatured, single-stranded (ss) DNA samples can be analyzed on urea–agarose gels. Here we report that after denaturation by heat in the presence of 8 M urea, the two strands of the same ds DNA fragment of ∼1–20-kb size migrate differently in 1 M urea containing agarose gels. The two strands are readily distinguished on Southern blots by ss-specific probes. The different migration of the two strands could be attributed to their different, base composition-dependent conformation impinging on the electrophoretic mobility of the ss molecules. This phenomenon can be exploited for the efficient preparation of strand-specific probes and for the separation of the complementary DNA strands for subsequent analysis, offering a new tool for various cell biological research areas. |
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