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

Éva Hegedüs, Endre Kókai, Alexander Kotlyar, V. Dombrádi, Gábor Szabó

Research output: Contribution to journalArticle

11 Citations (Scopus)

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
JournalNucleic Acids Research
Volume37
Issue number17
DOIs
Publication statusPublished - Jun 23 2009

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Agar Gel Electrophoresis
Urea
Complementary DNA
Sepharose
Gels
Single-Stranded DNA
Base Composition
Southern Blotting
Hot Temperature
DNA
Research

ASJC Scopus subject areas

  • Genetics

Cite this

Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis. / Hegedüs, Éva; Kókai, Endre; Kotlyar, Alexander; Dombrádi, V.; Szabó, Gábor.

In: Nucleic Acids Research, Vol. 37, No. 17, gkp539, 23.06.2009.

Research output: Contribution to journalArticle

Hegedüs, Éva ; Kókai, Endre ; Kotlyar, Alexander ; Dombrádi, V. ; Szabó, Gábor. / Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis. In: Nucleic Acids Research. 2009 ; Vol. 37, No. 17.
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