Synthesis and Purification in a Single Column on a High-Throughput Automated Oligonucleotide Production System

Abstract

A new oligonucleotide production system, the ABI 3948, was developed to automate the entire process: sequence entry, synthesis, cleavage, deprotection, purification, quantitation, sample collection and data management. Up to 48 primer-length, high-purity oligonucleotides can be produced on the ABI 3948 in a 24-h period of unattended operation. Synthesis, cleavage and purification all occur within the unique polystyrene-based OneStep® column. Oligonucleotides, analyzed by MicroGel™ capillary electrophoresis, anionexchange HPLC, polyacrylamide gel electrophoresis and enzymatic digestion/HPLC routinely demonstrate >90% product purity. Stringent PCR and automated fluorescent sequencing and genetic analysis experiments show the benefits of combined synthesis and purification of oligonucleotides.