Electrophoretic strand separation of long DNAs with poly (U,G) in agarose gels.
We have found that binding of poly(U,G) to single-stranded DNA decreases its mobility in 0.3% agarose gels. Differential binding to the complementary strands of denatured duplex DNA provides a simple method for strand separation. The method is shown to work with bacteriophage lambda DNA, adenovirus DNA and mtDNA for Tetrahymena pyriformis. In all cases the strand that binds more poly(U,G) in CsCl gradients also binds more in gels. The separated strands can be directly blotted from the gel onto nitrocellulose filters and used for hybridization experiments.
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002499:Centrifugation, Density Gradient / Q000379:methods; D003090:Coliphages / Q000235:genetics; D004272:DNA, Mitochondrial / Q000302:isolation & purification*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D009693:Nucleic Acid Hybridization; D011131:Polyribonucleotides; D013698:Templates, Genetic; D013769:Tetrahymena pyriformis / Q000235:genetics; D014158:Transcription, Genetic
D004272:DNA, Mitochondrial; D004279:DNA, Viral; D011131:Polyribonucleotides