Rapid and efficient oligonucleotide synthesis with low reagent consumption via a new synthesis column design: preparation of fluorescent dye labelled primers for application in PCR.

Abstract

A low dead-volume, 40 nmole scale column was designed for automated, solid-support oligonucleotide synthesis. The LV40 columns are filled with 1000A, high-cross link polystyrene beads at the 40 nmole scale. Reducing the unoccupied volume and optimizing the column dimensions allows efficient and fast synthesis on existing commercial synthesizers with low reagent consumption. Three spectrally distinct fluorescent dyes were applied as phosphoramidites in the synthesis of PCR primers. Fluorescent labelled PCR products of the Mfd11 microsatellite locus were analyzed.