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Oligonucleotide Purity Analysis by Capillary Electrophoresis


Introduction

Millions of oligonucleotides are synthesized each year for use as primers or probes in genetic analysis. Quality control of these preparations has traditionally been done by slab gel electrophoresis or HPLC, but these methods may require expensive reagents and laborious protocols, and may not provide desired quantitative precision or automation. Capillary electrophorsis (CE) using polymer solution buffers is being used increasingly for determination of oligonucleotide purity because of its high resolution, low consumption of sample and reagents, and ability to be fully automated. The dissolved polymers act as the sieving matrix and provide separations of oligonucleotides substantially equivalent to polyacrylamide and agarose gel electrophoresis.1, 2 This method is called dynamic sieving capillary electrophoresis (DSCE).

The CE Oligonucleotide Analysis Kit is optimized for use on the BioFocus capillary electrophoresis system. It can be used to determine the purity of synthetic oligonucleotides with quantitative estimation of the amounts of product and failure sequences. Separations are achieved by DSCE. The polymer type and concentration have been selected to provide unit resolution of oligonucleotides in the 440mer range. Replacement of the sieving buffer in the capillary between each analysis provides reproducibility of migration time and peak area. The analysis employs capillaries coated internally with a novel polymer (polyAAEE, acryloylaminoethoxyethanol) which ensures high-resolution separations over many runs.


CE Oligonucleotide Kit Components

The kit contains all the reagen
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