Fig. 4.
LC-QqQ-MS quantification of enzymatic cap modifications. For quantification, enzymatically modified RNAs were digested into nucleosides, leaving the cap either intact for Ecm1-catalyzed N7-methylation or digested for CAPAM-catalyzed N6-modification of Am in the TSN position. Samples were then analyzed via LC-QqQ-MS and quantified via external calibration. A) Formation of m7GpppG-FLuc RNA from GpppG-FLuc RNA by Ecm1 (100 mol%) and AdoMet. Unmodified mRNA is shown as control. Reproduced from Ref. [19] with permission from the Royal Society of Chemistry. B) Analysis of long mRNAs (RBD, eGFP and RLuc) after modification. RNAs were modified with CAPAM (200 mol%) using AdoMet or SeAdoYn. Unmodified mRNA is shown as control. Reproduced with permission from Ref. [20] licensed under CC BY-NC 4.0.