Synthesis of modified Nucleosides for use as Energy Transfer Probes in Genetic Analysis

Abstract

Energy transfer probes are used for target recognition in genetic analysis assays. The development of a novel energy transfer linker utilising a modified nucleoside (uridine or cytidine) is described.

The nucleoside (uridine or cytidine) was modified by the addition of an ethoxy functionality to the 2'-hydroxyl position. The synthesis of the phosphoramidites of both nucleosides is outlined.

The nucleosides were incorporated into oligonucleotides, as the phosphoramidites, and their performance as energy transfer linkers was investigated.

The energy transfer linker was incorporated into both Hybeacon and Taqman assays. the linker's ability to function as an energy transfer probe was assessed.

There is a good possibility of success with the Hybeacon assay as the energy transfer linker shows better discrimination between wild type and mutant sequences. Success with either of the above systems will lead to a commercially viable product.

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