Separation of (3′) Deoxynucleotides with Cation Exchange Columns

Abstract

A procedure is described for a one-step separation of synthetic (3′) deoxynucleotides purchased commercially or those purified and Isolated from the enzymatic digests of DNA. The method is simple. A 50 ml buret was used for column which was filled with the resin slurry and packed at water aspirator pressure. The compounds were eluted from the column at atmospheric pressure under gravity flow using a fraction collector and read on a Beck-man DU spectrophotometer. Recoveries were in excess of 90% and no accessory devices were required. The use of a volatile buffer, e. g. ammonium formate facilitated the recovery of the purified material by allowing the evaporation of the medium in which the sample was eluted.