Reversed-phase ion-pair liquid chromatography method for purification of duplex DNA with single base pair resolution.

ABSTRACT: Obtaining quantities of highly pure duplex DNA is a bottleneck in the biophysical analysis of protein-DNA complexes. In traditional DNA purification methods, the individual cognate DNA strands are purified separately before annealing to form DNA duplexes. This approach works well for palindromic sequences, in which top and bottom strands are identical and duplex formation is typically complete. However, in cases where the DNA is non-palindromic, excess of single-stranded DNA must be removed through additional purification steps to prevent it from interfering in further experiments. Here we describe and apply a novel reversed-phase ion-pair liquid chromatography purification method for double-stranded DNA ranging in lengths from 17 to 51 bp. Both palindromic and non-palindromic DNA can be r

SUBMITTER: Wysoczynski CL 

PROVIDER: S-EPMC3814375 | biostudies-literature | 2013 Nov

REPOSITORIES: biostudies-literature