GSM9965676: HCT116 spheroids, 5-fluorouracil 10 uM, rep 2; Homo sapiens; RNA-Seq
Spheroids were harvested by plate inversion (collection trays rinsed between replicates); the 128 wells per condition per plate were pooled in PBS, pelleted at 200 x g for 5 min and lysed. Total RNA was extracted with the RNAqueous-Micro kit (Thermo Fisher, AM1931) including on-column genomic DNA removal, eluted in IDTE buffer (10 mM Tris, 0.1 mM EDTA, pH 7.5; IDT) and quantified by NanoDrop (Thermo Fisher). RNA was stabilised with SEQguard Dino Preserve (Genovis, PS-SDP-024) for ambient-temperature shipping. Library preparation for 3'-end counting was performed by Plasmidsaurus using their standard commercial workflow. Libraries were sequenced on an Illumina NovaSeq X Plus to a depth of approximately 10 million deduplicated reads per sample.