lrRNA-Seq of Lung adenocarcinoma cell line
Total RNA was isolated from fresh cells using TRIzol Reagent (Invitrogen, 15596026). 500ng total RNA spiked in with Sequins (Garvan Institute) and SIRV-Set 4 (Lexogen, 141.01) was used to prepare libraries with the Oxford Nanopore PCR-cDNA Barcoding kit (SQK-PCB114.24). Reverse transcription was performed for 90 minutes at 42C. PCR amplification was performed with 14 cycles and an extension time of 4 minutes. Pooled barcoded libraries were loaded onto 3 PromethION R10.4.1 flowcells (FLO-PRO114M) at a concentration of 60fmol.