Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis

Éva Hegedüs, Endre Kókai, Alexander Kotlyar, Viktor Dombrádi, Gábor Szabó*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Article numbergkp539
Pages (from-to)e112-e112
JournalNucleic Acids Research
Volume37
Issue number17
DOIs
StatePublished - 23 Jun 2009

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