HEK-ADAR1-OEr2 - fast5
Total RNAs (totRNA) from S. cerevisiae (supplied by Prof N. Guaragnella at UNIBA), HEK293T cell lines (supplied by Dr. R. Pecori and Dr A. Arnold at DKFZ) as well as coli-IVTs poly(A)RNAs samples were used for Nanopore libraries preparation with the SQK-RNA002 - Direct RNA Sequencing Kit, following the protocol Direct RNA sequencing (SQK-RNA002) Version: DRS_9080_v2_revT_ 14Aug2019.For totRNA samples, the first two libraries were prepared from 500 ng of totRNA input, following the manufacturer's instructions. Subsequently, to increase the read number per sample, the totRNA input was increased up to 2 g. For poly(A) RNA samples, the first library was prepared from 50 ng of poly(A) RNA input; for the second library prep, the input was increased to 150 ng.Input RNA was concentrated using RNAClean XP (Beckman Coulter) beads 1.8X to reduce the sample volume, when necessary.Each library was loaded on a single R9.4.1 Promethion flow cell using the Flow Cell Priming Kit (EXP-FLP002), and the sequencing run was carried out on the P2 Solo device. Sequencing runs were monitored through the ONT MinKNOW software using the fast basecalling modality and stopped after about 72 hours. Raw reads in FAST5 format were processed for downstream analyses.