GSM7966853: CD14+ monocytes, 10 μM DG38:4, 2; Homo sapiens; RNA-Seq
Human monocytes were lysed using TriPure Isolation Reagent (Roche, Basel, Switzerland) and stored at -80 prior to RNA isolation. RNA was isolated according to the manufacturer's instructions, and the quality was determined using Lab-on-a-Chip RNA 6000 Nano on an Agilent Bioanalyzer (Agilent Technologies, Santa Clara, California, USA). Library construction, quality control and sequencing were performed at Novogene. In brief, messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional library or dTTP for non-directional library. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on a HiSeq instrument (Illumina) with 150-bp paired-end reads and 10 G data per sample. Obtained reads were processed through in-house perl scripts at Novogene and were aligned to the human genome using Hisat2 v2.0.5. Then reads were counted with featureCounts v1.5.0-p3.