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Nucleic Acids Research, 1978, Vol. 5, No. 8 2743-2754
© 1978

Articles

Electrophoretic strand separation of long DNAs with poly (U,G) in agarose gels

R.W. Goldbach+, R.F. Evers and P. Borst

Section for Medical Enzyrmology and Molecular Biology, Laboratory of Biochemistry, University of Amsterdam P.O. Box 60000, 1005 GA Amsterdam, Netherlands

Received June 8, 1978.

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 complement ary 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.

+Present address: Laboratory of Molecular Biology, Agricultural University, De Dreijen 11, Wageningen, The Netherlands


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