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. 2014 Jun 30;42(15):e117. doi: 10.1093/nar/gku553

Figure 2.

Figure 2.

Synthesis of site-specific spin-labeled RNA. (A) Native gel electrophoresis on 10% polyacrylamide. RNA segments and DNA splint were loaded in the T4 RNA ligase 2 buffer: lane 1, (pG246T-C55); lane 2, (pppG1-G23); lane 3, DNA splint; lane 4, (pppG1-G23) + (pG246T-C55) + DNA splint. (B) Denaturing 12% polyacrylamide gel. Lane 1: RNA fragment (G1-G23) acceptor; lane 2: RNA fragment (pG246T-C55) donor; lane 3: DNA splint (43-nt); lane 4: preparative ligation; lane 5: purified ligation product. (C) Imino-proton region of 1D spectra recorded at 20°C of the wild-type RNA full-length (top) and the ligation product (bottom). All imino protons were assigned via sequential Nuclear Overhauser Effects (NOEs) observed in 2D-NOESY experiments, with the exception of the resonances at 10.37, 11.09 and 11.43 p.p.m. that could not be identified unambiguously.