The use of multiple liquid chromatography methods augmented by phosphorus-31 nuclear magnetic resonance to characterize the diastereomer composition in synthetic oligonucleotides

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P NMR provided complementary and additional insight into the diastereomer composition of phosphorothioate-linked oligonucleotides. Further, our data also indicated that the chemical activator substantially influenced the diastereomer content but other important factors including the overall chemical synthesis process and sequence specific nucleobases and modifications play an important role as well.