Abstract
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.
MeSH Terms
Adenoviridae/genetics
Animals
Centrifugation, Density Gradient/methods
Coliphages/genetics
DNA, Mitochondrial/isolation & purification
DNA, Viral/genetics,isolation & purification
Electrophoresis, Agar Gel/methods
Nucleic Acid Hybridization
Polyribonucleotides
Templates, Genetic
Tetrahymena pyriformis/genetics
Transcription, Genetic
Chemicals
DNA, Mitochondrial
DNA, Viral
Polyribonucleotides
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Goldbach R W
Evers R F
Borst P
References (17)
17 references, click to expand
-
Cleavage maps of DNA from adenovirus types 2 and 5 by restriction endonucleases EcoRI and HpaI.
Cold Spring Harb Symp Quant Biol. 1975;39 Pt 1:397-400
PMID: 1057470
-
Detection of specific sequences among DNA fragments separated by gel electrophoresis.
J Mol Biol. 1975 Nov 5;98(3):503-17
PMID: 1195397
-
The binding of poly(rA) and poly(rU) to denatured DNA. I. Model studies with homopolymers.
Nucleic Acids Res. 1976 Apr;3(4):1013-27
PMID: 1272800
-
The binding of poly (rA) and poly (rU) to denatured DNA. II. Studies with natural DNAs.
Nucleic Acids Res. 1976 Apr;3(4):1029-51
PMID: 1272801
-
COHESION OF DNA MOLECULES ISOLATED FROM PHAGE LAMBDA.
Proc Natl Acad Sci U S A. 1963 May;49(5):748-55
PMID: 16591099
-
Preparative separation of the complementary strands of DNA restriction fragments by alkaline RPC-5 chromatography.
Nucleic Acids Res. 1978 Jan;5(1):13-21
PMID: 25414
-
Separation of the complementary strands of DNA fragments on polyacrylamide gels.
Nucleic Acids Res. 1977;4(5):1569-78
PMID: 331258
-
The structure of Tetrahymena pyriformis mitochondrial DNA. II. The complex structure of strain GL mitochondrial DNA.
Biochim Biophys Acta. 1977 Jul 5;477(1):51-69
PMID: 406927
-
Transcription of Tetrahymena mitochondrial DNA in vivo.
Biochim Biophys Acta. 1973 May 18;308(3):372-80
PMID: 4197078
-
Patterns of interaction between polyribonucleotides and individual DNA strands derived from several vertebrates, bacteria and bacteriophages.
J Mol Biol. 1966 Sep;20(2):313-29
PMID: 4291221
-
Gel electrophoretic separation of the complementary strands of bacteriophage DNA.
Virology. 1972 Jul;49(1):342-4
PMID: 5039027
-
The binding of polyribonucleotides to the complementary strands of mitochondrial DNA.
Biochim Biophys Acta. 1972 Feb 15;259(3):313-20
PMID: 5062502
-
The gel electrophoresis of DNA.
Biochim Biophys Acta. 1972 May 10;269(2):192-200
PMID: 5063906
-
Effect of size and aggregation of guanine-rich polyribonucleotides on the practical aspects of DNA strand separation in CsCl density gradients.
Biochim Biophys Acta. 1969 May 20;182(1):269-72
PMID: 5792856
-
Amplification and characterization of a beta-globin gene synthesized in vitro.
Cell. 1976 Jun;8(2):163-82
PMID: 61066
-
The organization of genes in yeast mitochondrial DNA II. The physical map of EcoRI and HindII + III fragments.
Mol Gen Genet. 1975 Dec 30;143(1):53-64
PMID: 765743
-
Adenovirus transcription. III. Mapping of viral RNA sequences in cells productively infected by adenovirus type 5.
Virology. 1976 Jul 15;72(2):443-55
PMID: 948869