GSM9293394: rCD4_NMS_24h; Homo sapiens; RNA-Seq
Total RNA was extracted from all samples using TRIzol reagent (Thermo Fisher Scientific, USA) following the manufacturer's instructions. RNA concentration and purity were measured using a NanoDrop spectrophotometer (Thermo Fisher Scientific, USA), and RNA integrity was assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies, USA). Only RNA samples with an RNA integrity number (RIN) > 7 were used for downstream library preparation. Comprehensive RNA libraries, including long non-coding RNAs and mRNAs, were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs, USA). Ribosomal RNA was removed from total RNA using the Ribo-Zero rRNA Removal Kit (Illumina, USA). The rRNA-depleted RNA was fragmented and reverse-transcribed into cDNA, followed by end repair, adaptor ligation, and PCR amplification according to the manufacturer's protocol. The resulting libraries were purified using AMPure XP beads (Beckman Coulter, USA). Library quality was assessed using an Agilent 2100 Bioanalyzer, and concentrations were measured by qPCR. Sequencing was performed on the DNBSEQ-T7 platform generating 150-bp paired-end reads.