DNA covalently linked to carboxymethyl-cellulose and its application in affinity chromatography.

Affinity chromatography on columns containing DNA has been successfully used in the isolation and purification of DNA dependent polymerases and other DNA-binding proteins as well as for the purification of nucleic acids hybridizable to the column bound DNA [l] . Two drawbacks of DNA column materials used so far have limited their application: firstly, in the most widely used column materials DNA-cellulose [2] and DNA-agarose [3] the nucleic acid is not bound covalently, imposing a restriction of the ionic strength of the buffers used to avoid a loss of DNA; secondly, in those instances where DNA was bound covalently to the matrix, the amount of bound DNA was very low [4,5]. We have therefore been interested in developing a column material containing covalently linked DNA in amounts comparable to those in DNA-agarose. In this paper we describe the preparation of an affinity chromatography material containing up to 3.6 mg DNA per ml bedvolume and its successful application in the purification of DNA dependent DNA polymerases from yeast nuclei and mitochondria. While this work was in progress Arndt-Jovin et al. [6] reported an improved method for the preparation of agarose containing similarly high amounts of covalently attached DNA.