Covalently Mercurated Molecular Beacon for Discriminating the Canonical Nucleobases

A highly nucleobase-discriminating metalated nucleoside analogue, 3-fluoro-2-mercuri-6-methylaniline, was incorporated into an oligonucleotide molecular beacon. Fluorescence emission spectra were measured after the addition of four different complementary strands, in which the nucleobase opposite the metalated analogue varies. The fluorescence results showed a clear binding selectivity at room temperature, in the order G>T>C>A. The selectivity is based on the different affinities between the metalated nucleoside analogue and the canonical nucleobases. The synthesized probe is capable of robust discrimination between the two purine as well as the two pyrimidine bases by fluorescence at room temperature, and more sophisticated temperature analysis allows clear separation of every canonical nucleobase. The probe would, hence, be a suitable method for the detection of single nucleotide polymorphisms.

Automated summary

By incorporating a highly nucleobase-discriminating metalated nucleoside analogue into an oligonucleotide molecular beacon, clear binding selectivity was achieved at room temperature, with a preference order of G>T>C>A. This method allows for robust discrimination between different nucleobases and is suitable for the detection of single nucleotide polymorphisms.