Skip Navigation

This Article
Right arrow Print PDF (1838K)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Zuidema, D.
Right arrow Articles by Flavell, R.A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zuidema, D.
Right arrow Articles by Flavell, R.A.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1978, Vol. 5, No. 7 2471-2483
© 1978

Articles

The isolation of duplex DNA fragments containing (dG.dC) clusters by chromatography on poly(rC)-Sephadex

D. Zuidema, F.M. van den Berg and R.A. Flavell

Section for Medical Enzymology and Molecular Biology, Laboratory of Biochemistry, University of Amsterdam PO Box 60.000, 1005 GA Amsterdam, Netherlands

Received March 20, 1978. Duplex DNA containing oligo(dG.dC)-rich clusters can be isolated by spe cific binding to poly(rC)-Sephadex. This binding, probably mediated by the formation of an oligo(dG.dC)rC+ triple helix, is optimal at pH 5 in 50% for mamide, 2 N LiCl; the bound DNA is recovered by elution at pH 7.5.

Using this method we find that the viral DNAs PM2, lambda and SV40 con tain at least 1, 1 and 2 sites for binding to poly(rC)-Sephadex, respectively. These binding sites have been mapped in the case of SV40; the binding sites can in turn be used for physical mapping studies of DNAs containing (dG.dC) clusters. Inspection of the sequence of the bound fragments of SV40 DNA shows that a (dG.dC) tract is required for the binding of duplex DNA to poly(rC)-Sephadex. Although about 60% of rabbit DNA cleaved with restric tion endonuclease KpnI binds to poly(rC)-Sephadex, no binding is observed for the 5.1 kb DNA fragment generated by KpnI digestion, which contains the rabbit ß-globin gene. This indicates that oligo(dG.dC) clusters are not found close to the rabbit ß-globin gene.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.