HPLC analysis of synthetic oligonucleotides using spectral contrast techniques

The need for high-purity oligodeoxyribonucleotides for various applications has resulted in the development of novel synthesis, purification and analytical techniques. A diversity of methods, including polyacrylamide slab gel electrophoresis, capillary gel electrophoresis, as well as HPLC, have been successfully used to analyze material throughout the purification process. This study demonstrates how the application of spectral comparison techniques to synthetic products resolved by anion-exchange HPLC can distinguish deletion fragments (i.e., "N-1" sequences) from full-length products. Such analysis techniques can also differentiate partial from fully phosphorothioated DNA sequences. In combination, HPLC separation and spectral analysis technology provide information not obtainable with any other single analytical method.