RAPID SCALEABLE CHROMATOGRAPHIC PURIFICATION OF NUCLEIC-ACIDS FROM PROTEINACEOUS MIXTURES

The present studies describe a unique HPLC method for the purification of nucleic acids from proteins, scaleable to a preparative or process level, using a Zorbax PRO-10/DN 300 Angstrom silica packing. Columns of this material have an extremely high affinity for proteinaceous materials, and a very low affinity for nucleic acids. We demonstrate this by separating salmon DNA from bovine serum albumin; and describe use of the PRO-10/D-N column to separate plasmid DNA from bacterial lysates. The method is quick and, unlike phenol-chloroform extraction, utilizes physiological buffers to minimize hazardous operations. This LC method is faster than classical non-LC methods, is less labor intensive, and uses no organic solvents, thereby eliminating the need for post-separation recovery operations. The high recovery of DNA purified by Zorbax PRO-10/D-N provides DNA material suitable for many common molecular manipulations including restriction enzyme cleavage, ligation, and amplification.