RNAseq_RUES2-1st-D29-1-3
Cells were dissociated using TrypLE Select, and resuspended in FACS buffer to stop digestion. Cells were pelleted, frozen at -80 °C temporarily, and lysed with TRIzol Reagent (Thermo Fisher Scientific). RNA was extracted using Direct-zol RNA Miniprep Plus kit (Zymo Research). RNA concentration and quality was measured by 2100 Bioanalyzer (Agilent Technologies). Libraries were prepared using SMART-Seq v4 Ultra Low Input RNA kit (Takara Bio) and sequenced by NovaSeq (Illumina) with paired-end 100 bp reads at MedGenome. Each library was sequenced with a targeted depth of 40 million total reads.