GSM7225337
in vitro transcribed RNA was first synthesized and PAGE purified prior to modification with 2A3 and then reverse transcribed. Resulting cDNA was used as input material for library assembly. RNA is first modified with 2A3 or treated with DMSO control, then fragmented to a median length of 150nt. The fragmented material is used as input for a random hexamer reverse transcription reaction, afterwhich the resulting cDNA is used as starting material for NEBNext Ultra II DNA library prep kit for Illumina. 2'-hydroxyl acylation analyzed by primer extension (SHAPE) Mutational Profiling via High Throughput Sequencing (MaP-seq).