Extended Data Fig. 2: Application of hybrid solid-liquid phase PORDVA in synthesizing RNA with multiple modifications at discrete positions.
From: Engineering a DNA polymerase for modifying large RNA at specific positions
(a) Synthesis scheme for P14 with an internal m7G (at residue 26) and Cy3 (at residue 39) using the hybrid-phase PORDVA. (b) Images of denaturing PAGE of 10 µL reaction mixture collected from steps 1 to 4 after treatment with DNase I and proteinase K at 37 °C for 30 min. 25 nt LRNA and 45 nt unmodified RNA were loaded into Lanes 1 and 6, respectively, as controls. (c) LC-MS of P14. The DNA and RNA sequences used in the hybrid-phase PORDVA synthesis are listed in Supplementary Table 21.
