GSM7051360: pre-treatment blood14; Homo sapiens; RNA-Seq
Buffycoat or PBMCs were isolated from whole blood. Total RNA was isolated using Trizol (Invitrogen, Waltham, MA, USA) and the purity and concentration were verified using a NanoDrop ND-1000 instrument (ThermoFisher Scientific, Waltham, MA, USA). The integrity of the RNA was assessed by a 2100 Bioanalyzer gel image analysis system (Agilent, Santa Clara, CA, USA). Only RNA samples with an integrity number of 7 or higher were processed for RNA-seq. At least 100 ng mRNA per sample was submitted for library preparation. Qualified RNA samples were enriched and synthesized into cDNA for library preparation. RNA-seq libraries were constructed using the TruSeq RNA Sample Prep Kit. The libraries from qualified RNA samples were sequenced in the 150 nt paired-end mode on an Illumina NovaSeq 6000 platform. After quality filtering, over 30 billion clean reads were obtained in each library and then used for down-stream analysis.