title: HeLa nucleoli ONT
design: Total RNA from isolated HeLa nucleoli was depleted of rRNA by the RiboMinus Human/Mouse Transcriptome Isolation Kit (Invitrogen). RNA was converted to double stranded cDNA using random hexamer with the NEBNext Ultra RNA First Strand and NEBNext Ultra RNA 2nd Strand Synthesis Kits (NEB). 1D library was prepared with the SQK-LSK108 kit (Oxford Nanopore) and sequenced on a SpotON Flowcell MK I (R9.4) flowcell for 14 h using a MinION MK 1B sequencer. The flowcells was washed and another identical library was loaded in the same flowcell and sequenced for another 14 h. Basecalling was performed with Albacore 2.0.2