title: MinION sequencing; Mus musculus bone marrow-derived macrophages, TENT5A Flox/Flox TENT5C KO, direct RNA sequencing, biological replicate 2
library_construction_protocol: Total BMDM RNA was isolated with TRI Reagent according to the manufacturer's instructions (Sigma-Aldrich). Nanopore Direct RNA libraries were prepared using Direct RNA Sequencing KIT SQK-RNA002 (ONT) according to the manufacturer's instructions, from 3.5 μg of cap-enriched mRNA mixed with 150 ng of Saccharomyces cerevisiae oligo(dT)-enriched mRNA to optimize sequencing efficiency. Sequencing was performed with a MinION device (ONT; Flow cell type FLO-MIN106, RevD). Raw reads were basecalled with the standalone version of Guppy 4.0.11 (ONT).