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A) N2-double methylation by hTgs1 of a 7mG-capped RNA (4) with 6mAm as the first nucleotide yielding 7. B) RP-HPLC analysis of double N2-methylation reaction of 7mGppp 21-mer 4 after 7.5 h incubation with hTgs1 at 2.5 μM (black trace) in comparison with the N7-methylated substrate 4 (red trace). C) MALDI-TOF mass spectrum (right panel) of the trimethylated capped 21-mers 2m,2m,7mGppp6mAmCACUUGCUUUUGACACAACU 7.
