A capture approach for supercoiled plasmid DNA using a triplex-forming oligonucleotide.

ABSTRACT: Proteins that recognize and bind specific sites in DNA are essential for regulation of numerous biological functions. Such proteins often require a negative supercoiled DNA topology to function correctly. In current research, short linear DNA is often used to study DNA-protein interactions. Although linear DNA can easily be modified, for capture on a surface, its relaxed topology does not accurately resemble the natural situation in which DNA is generally negatively supercoiled. Moreover, specific binding sequences are flanked by large stretches of non-target sequence in vivo. Here, we present a straightforward method for capturing negatively supercoiled plasmid DNA on a streptavidin surface. It relies on the formation of a temporary parallel triplex, using a triple helix forming oligonucl

SUBMITTER: Ruigrok VJ 

PROVIDER: S-EPMC3664820 | biostudies-literature | 2013 May

REPOSITORIES: biostudies-literature