Anion Exchange Purification of DNA

Hamilton PRP-X600 anion exchange HPLC columns separate DNA oligomers according to charge. Controlled porosity provides excellent separation of n-1 oligomers from the full length product. See application #11 for the separation of a DMT-on antisense, phosphorothioate 18mer from the failure sequences. PRP-X600 columns work best when heated to 85°C.

DNA Sample Capacity

The 50 mm long 4.6 mm I.D. columns have a sample capacity of approximately 1 mg.150 x 4.6 mm long columns have a sample capacity of about 3 mg.

Compatible Mobile Phases for DNA Purification

PRP-X600 weak base anion exchange columns are polymeric, stable from pH 1 to 11 and compatible with organic solvents.

  • 20 mM TRIS, 1 mM EDTA
  • 1N Sodium Chloride in 20 mM TRIS, 1 mM EDTA
  • 85:15 100 mM TRIS pH 8.0 : Acetonitrile
  • 85:15 100 mM TRIS pH 4.0,2.5 M Lithium Chloride
  • Acetonitrile

Changing Mobile Phases and Equilibration After Gradient Separation

Equilibration of the PRP-X600 column requires approximately 10 column volumes. Following each gradient analysis, reequilibrate the column with Buffer A to ensure reproducible sample retention.

PRP-X600, 4.6 x 50 mm (2.0 mL/min)

DMT-on Phosphorothioate Antisense 18mer