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Abstract
In this technical note, we systematically describe the chemical synthesis, deprotection, and purification of diphosphorylated RNA oligonucleotides. This process eliminates the need for an enzymatic step after chemical synthesis of a 5’triphosphate RNA, allowing the user to synthesize the 5’diphosphate RNA directly. This protocol can also be applied to other chemically synthesized nucleic acids, including those with modifications.
