GSM7890428: Mild_1; Homo sapiens; RNA-Seq
Blood was collected in Vacutainer EDTA tubes (BD Biosciences). Human neutrophils were isolated using MACSxpress® Whole Blood Neutrophil Isolation Kit, human (Miltenyi Biotech) following the manufacturer's protocol. Purity of isolated neutrophils was assessed using flow cytometry (CD15+CD16+CD14-CD193-). Greater than 90% purity was achieved for all samples. RNA was isolated using RNeasy Mini kit (Qiagen) and RNA integrity was analyzed by 2100 Expert Bioanalyzer System (Agilent), using the Agilent RNA 6000 Pico Kit (Agilent). Libraries were simultaneously prepared from extracted total RNA using Illumina Truseq Stranded mRNA library preparation kit according to the manufacturer's protocol (Illumina). Prepared libraries were sequenced on the Illumina Nextseq500 at 30 million reads per sample at 2x75 cycles.