lrRNA-Seq of small cell lung cancer 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.