GSM5668944: NSTEMI_3_B; Homo sapiens; RNA-Seq
Human peripheral blood neutrophils were isolated by diluting peripheral venous blood collected in to EDTA tubes 1:1 with PBS and neutrophils were negatively enriched using the EasySep direct human neutrophil isolation kit as per the manufacturers instructions. Isolated neutrophils were with lysed with RLT (Qiagen) and stored at -80ºC. RNA was isolated using RNeasy Mini Kit (Qiagen), according to manufacturer's instructions. RNA-seq libraries were prepared with the TruSeq Stranded mRNA LT sample preparation kit (Illumina) using Sciclone and Zephyr liquid handling robotics (PerkinElmer). Library amounts were quantified using the Qubit 2.0 Fluorometric Quantitation system (Life Technologies) and the size distribution was assessed using the Experion Automated Electrophoresis System (Bio-Rad). For sequencing, libraries were pooled, diluted and sequenced on an Illumina HiSeq 3000 instrument using 50 bp single-read chemistry.