FAQ: Is it possible to use NudC to generate ligatible 5′ monophosphate RNA from non-canonical initiating nucleotides only?

For applications where non-canonical cap structures are to be distinguished from G-capped RNA in a 5′ ligation-based workflow, one can pre-treat the RNA sample with yDcpS (NEB #M0463), followed by Quick CIP (NEB #M0525) and a RNA purification step to turn G-capped RNA and existing 5′ phosphorylated RNA to 5′hydroxyl RNA, which is not available for ligation.