The DNAPac® PA200 provides the highest resolution of any commercially available column on the market. Like the DNAPac PA100, this column is stable under denaturing conditions, and is both rugged and reliable. However, it surpasses even the DNAPac PA100 with its ability to resolve the full-length product from its failure sequences and its stability under alkaline conditions. N, N – 1 resolution for oligonucleotides Resolve oligonucleotides with secondary structures Compatible with solvent, high pH, and high temperatures Retention control by eluent pH Selectivity control with eluent pH, salt, and solvent
Chromatographic oligonucleotide analyses, either by reversed-phase or by anion-exchange chromatography, are used for a variety of applications, such as to evaluate coupling efficiency of a synthesis, assess purity, and verify the nature of components in a nucleic acid amplification mixture. Strong anion-exchange chromatography, using pH-stable polymeric resins, is a simple and effective technique for separating oligonucleotides. One advantage of anion-exchange chromatography over reversed-phase chromatography using polymer-based materials is the ability to operate at high pH. Denaturing conditions established at high pH can be used to eliminate hydrogen bonding and allow resolution of problem sequences, such as self-complimentary sequences or poly-G stretches. As a consequence, anion-exchange chromatography at high pH has become the preferred approach for oligonucleotide analyses. The DNAPac PA200 is a strong anion-exchange column developed to provide unsurpassed high-resolution analysis and purification of synthetic oligonucleotides. By building on the unique features that have made the DNAPac PA100 the ultimate in nucleic acid separations, and incorporating additional features into the packing material, the DNAPac PA200 allows resolution of full length from n – 1, n + 1, and other failure sequences not possible with other columns. Retention times and selectivity can be modified, depending upon the choice of salt, pH, and solvent. Therefore, the separation can be easily tailored to the requirement of your specific analytical problem. The DNAPac PA200 is packed with a pellicular anion-exchange resin comprised of a 8-µm-diameter nonporous polymeric substrate to which is bound quaternary amine-functionalized MicroBeads. The rapid mass transport characteristics of this resin result in higher-resolution oligonucleotide separations than are possible with traditional macroporous resins or reversed-phase columns. The DNAPac PA200 can be operated under denaturing conditions, such as high temperature (up to 85 °C), high-pH eluents (up to pH 12.5), or with the inclusion of chaotropic agents. However, if the goal is to scale up to semipreparative separations, the DNAPac PA100 should be used, due to its higher capacity.
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