Ultrahigh-Resolution Separations of Oligonucleotides

The DNAPac^® PA200 is a strong anion-exchange column developed to provide unsurpassed high-resolution analysis and purification of synthetic oligonucleotides. DNAPac PA200 can resolve full length from n–1, n+1, and other failure sequences not possible with other columns. Retention times and selectivity can be controlled by the choice of salt, pH, and solvent. Therefore, the separation can be tailored to the requirements of many different oligonucleotide analysis challenges. 

• Achieve n, n-1 resolution for oligonucleotides
• Resolve oligonucleotides with secondary structures
• Assay phosphorothioate purity
• Selectivity control with eluent pH, salt, and solvent
• Resolve RNA with aberrant (2’–5’) links from normal SSRNA

The DNAPac PA200 is packed with a pellicular anion-exchange resin composed of an 8-μm-diameter nonporous polymeric substrate, to which is bound quaternary amine-functionalized MicroBeads^™. The rapid mass transport characteristics of this resin result in high-resolution oligonucleotide separations. 

The DNAPac PA200 can be operated under denaturing conditions, such as high temperature (up to 85 °C), or high-pH eluents (up to pH 12.5), or by the inclusion of chaotropic agents (such as Urea).

The DNAPac PA200 offers the highest quality phase stability over a broad pH range, delivers exceptional resolution and supports high-throughput separations. DNAPac PA200 columns support resolution of normal-length oligonucleotides (8- to 30- mer), extended-length oligonucleotides (30- to 70-mer), linear double-stranded DNA, and supercoiled versus nicked/linear DNA.

Analytical separations on the 4 mm diameter column can be scaled directly to larger diameter columns so preparative methods can be conveniently developed using small samples. Scaling the flow rate and sample size up for the 9 × 250 mm and 22 × 250 mm column yields essentially identical chromatography.