SHAPE-Seq of telomerase RNA Physcomitrium Patens
10 pmol of in vitro transcribed RNA was denatured at 95 C for 2 minutes, quickly cooled down on the ice, mixed with a folding buffer (333 mM HEPES, pH 8.0; 333 mM NaCl; 33 mM MgCl2) and folded at 37 C for 20 minutes. This RNA was used for the modification reactions with 100 mM 1M7 (Sigma Aldrich) (+), DMSO (-), and DC (denaturing control). Modified RNAs prepared in triplicates were purified with SPRI beads and used for reverse transcription with Superscript-II in the presence of MnCl2 (SHAPE-Seq).