Extended Data Fig. 1: Applications of hybrid solid-liquid phase PORDVA in synthesizing RNA with 5′-proximal and internal modifications.
From: Engineering a DNA polymerase for modifying large RNA at specific positions
(a) Synthesis schemes for P10 and P11 with Cy3 and 2′-OMe, respectively, at the 5′-end using the hybrid-phase PORDVA. (b) Secondary structures of P10-P13 synthesized via the hybrid-phase PORDVA. (c) Synthesis schemes for P12 and P13 labeled with internal m6A and m7G, respectively, using the hybrid-phase PORDVA. (d) Denaturing PAGE images of P10-P13 (Lanes 1 to 4) under UV and fluorescence excitation. The 45 nt unmodified RNA was loaded to Lane 5 as a control. (e) MS spectra of P10, P11, P12, and P13. (f) Native agarose gel image of DNA cleavage by Cas12a in the presence of unmodified RNA (Lane 2), P10 (Lane 3), P11 (Lane 4), P12 (Lane 5) and P13 (Lane 6). The target DNA was loaded to Lane 1 as a control. The DNA and RNA sequences used in the hybrid-phase PORDVA synthesis are listed in Supplementary Table 21.
