For n, n-1 Resolution of Single-Stranded Oligonucleotides

The DNAPac^®PA100 is a high-resolution anion-exchange column that provides single-base resolution. It is stable under denaturing conditions, rugged, reliable, and can be readily scaled up. The DNAPac PA100 is a 13 μm pellicular, nonporous polymeric resin with bound quaternary amine-functionalized MicroBeads^™. The rapid mass-transport characteristics of this resin result in very high-resolution oligonucleotide separations.

• Capable of n, n-1 resolution for oligonucleotides

• Resolves oligonucleotides with secondary structures

• Compatible with solvent, high pH, and high temperatures

• Analyzes phosphorothioate-based clinical samples

• Provides easy scale-up

The MicroBead resin used in DNAPac columns provides high-resolution separations to ensure maximum peak purity, and makes linear scale-up of your separation easy. For more complex separations, the polymeric properties of the DNAPac PA100 allows great flexibility to select ideal conditions, including high pH, high temperature, or chaotropic agents.

Synthetic oligonucleotides can be screened for production yield and failure sequences using the DNAPac PA100. Unit-base resolution of synthetic oligonucleotides to 60 bases and beyond has been demonstrated. The DNAPac PA100 can even be operated under denaturing conditions. Either high temperature (up to 90 °C) or high-pH eluents (up to pH 12.5) can be used to eliminate hydrogen bonding, allowing resolution of problem sequences such as self-complementary sequences or poly-G stretches.

Separates RNA / Double-Stranded DNA

The DNAPac PA100 is ideally suited for the purification and analysis of synthetic RNA. Failure sequences are easily separated from the full-length product. Double-stranded DNA, such as plasmids, or restriction fragments are also separated.

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. The loading capacity of the DNAPac PA100 (4 × 250 mm) column is from 1 to 100 μg. Scaling the flow rate and sample size up for the 9 × 250 mm and 22 × 250 mm column yields essentially identical chromatography.

Separation of oligonucleotides using the DNAPac^®PA100.

(A) Attempted Separation of 3 μg of pd(G)_12-18with a NaCl gradient of pH 8.0. (B) Resolution of pd(G)_12-18Homopolymers at pH 12.4 with a 500 to 900 mM NaCl Gradient.

Analysis of Synthetic Homopolymer.