GSM8619009: Basophils_4; Homo sapiens; RNA-Seq
RNA was isolated from sorted basophils (the in vitro testing) using the RNeasy Plus Mini Kit (Qiagen, Hilden, Germany) and from PAX RNA tubes (the clinical cohort) using the PAXgene Blood miRNA Kit (PreAnalytiX, Hombrechtikon, Switzerland), following the manufacturers' recommendations. RNA concentration was determined using the Synergy2 microplate reader (Biotek, Winooski, VT, USA), and RNA integrity was assessed using the Bioanalyzer 2100 (Agilent Technologies, Waldbronn, Germany) or by gel electrophoresis. For RNA sequencing (RNAseq), cDNA libraries were prepared using the Stranded mRNA Prep Ligation Kit (Illumina, San Diego, CA, USA) according to the manufacturer's instructions. Paired-end RNA sequencing (2×74 bp) was performed on a NextSeq 550 system (Illumina) using a NextSeq 500/550 Mid Output Kit v2.5 (Illumina).