GSM8969651: iPSC HELLS KO rep3 RNA; Homo sapiens; RNA-Seq
iPSCs were harvested using a single-cell suspension (see growth protocol) and snap-frozen for further use. RNA was extracted using the RNeasy Plus Micro Kit (Qiagen, #74034). Three biological replicates were collected per condition. Each sample was resuspend-ed in 350µl of RLT buffer containing 10% β-mercaptoethanol (Thermo, #21985023), followed by the addition of 350µl of 70% ethanol. RNA was then purified according to the manufacturer's in-structions. RNA quality and concentration were assessed prior to library preparation. For RNA sequencing, the KAPA RNA HyperPrep Kit (Roche, # KK8560) was used to generate libraries from 500ng RNA per sample. The final libraries were purified using Agencourt AMPure XP beads, and diluted to 4nM for 100 bp paired-end sequencing on Element Biosciences' AVITI system.