GSM9032760: U937 cells, non-template control, rep 2; Homo sapiens; RNA-Seq
Total RNA was isolated using the Direct-zol RNA Miniprep Plus kit (R2072, Zymo Research) 500 nanograms of total RNA were used for NEBNext Ultra II Directional RNA Library Prep Kit (NEB # E7760L). Poly-A selection (NEBNext Poly (A) Magnetic Isolation Module; NEB # E7490) and cDNA synthesis were performed according to NEB protocol. The NEBNext UMI RNA Adaptors were diluted 50-fold in ice-cold UMI Adaptor dilution buffer. Ligation reaction was purified using Ampure XP beads. (Beckman Coulter, # A63881) and the adaptor ligated cDNA was amplified with 12 cycles of PCR with NEBNext Primer Mix (96 Unique Dual Index UMI Adaptors RNA set1, (NEB #E7416). Equimolar amounts of the samples were then pooled and sequenced using NextSeq 500/550 70 base read lengths in paired-end mode.