GSM9293404: rCD4_GW_48h; Homo sapiens; miRNA-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. Small RNA libraries were prepared using the NEBNext Small RNA Library Prep Set for Illumina (New England Biolabs, USA) according to the manufacturer's protocol. Briefly, 3' and 5' adaptors were ligated to purified small RNAs, followed by first-strand cDNA synthesis and PCR amplification. The amplified libraries were purified using AMPure XP beads (Beckman Coulter, USA). Library quality and fragment size distribution were examined using an Agilent 2100 Bioanalyzer, and library concentration was quantified by qPCR. Sequencing was performed on the DNBSEQ-T7 platform generating 150-bp paired-end reads.