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A synthetic protocol for  34 S‐labeled phosphorothioate oligonucleotides (PS ONs) was developed to facilitate MS‐based assay analysis. This was enabled by a highly efficient, two‐step, one‐pot synthesis of  34 S‐labeled phenylacetyl disulfide ( 34 S‐PADS), starting from  34 S‐enriched elemental sulfur ( 34 S 8 ).  34 S‐PADS was subsequently used for stable isotope labeling (SIL) of oligonucleotides containing a phosphorothioate backbone. The  34 S‐SIL PS ONs are shown to retain the same melting temperature, antisense activity, and secondary structure as those of the corresponding unlabeled  32 S PS ONs.

A Versatile and Convenient Synthesis of 34 S‐Labeled Phosphorothioate Oligonucleotides