title: GSM8842352: Reverse complement, in vitro SHAPE experiment, 5NIA-treated, replicate 2; Homo sapiens; OTHER
library_construction_protocol: For in-cell probing, RNA was isolated with Trizol reagent. For in vitro probing, RNA was generated by in vitro transcription with NEB HiScribe T7 High Yield RNA Synthesis kit For MaP RT, superscript II was used to generate cDNA. For JuMP, C8 enzyme was used to perform RT. In-cell and SHAPE-JuMP libraries were constructed with two rounds of PCR to add Illumina adaptor sequences and barcodes. In vitro MaP libraries were prepared using the NEBNext Ultra II FS DNA Library Prep kit for Illumina