Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis.
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 approximately 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.
Hegedüs
Eva
E
Department of Biophysics and Cell Biology, University of Debrecen, 4012 Debrecen, Nagyerdei krt. 98, Hungary.
Kókai
Endre
E
Kotlyar
Alexander
A
Dombrádi
Viktor
V
Szabó
Gábor
G
D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000737:chemistry / Q000302:isolation & purification; D004277:DNA, Single-Stranded / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D009691:Nucleic Acid Denaturation; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014508:Urea / Q000737:chemistry*
D018076:DNA, Complementary; D004271:DNA, Fungal; D004277:DNA, Single-Stranded; D014508:Urea