GSM5025821: 029_CC; Homo sapiens; RNA-Seq
Peripheral blood was collected from patients using standard venipuncture techniques into PreAnalytiX PAXgene blood DNA and RNA tubes. Vacutainers were inverted several times and stored at -80°C. Samples were isolated from peripheral leukocytes according to manufacturer instructions with the PAXgene DNA or RNA isolation kits (Qiagen). Isolated samples were stored at -20°C. mRNA was sequenced using the Illumina TruSeq RNA Library Prep kit on the HiSeq 2000 platform following the manufacturer's protocol. Paired end RNA reads were aligned to human genome reference (GRCh37 ensembl version 70), using STAR RNAseq read aligner (33), and accepted mapped reads were summarized to gene level counts using the “featureCounts” function of the subread software package (34). Differentially expressed genes (DEGs) were reported by the DESeq2 package (35) from the count matrix following standard protocol, using age + sex + RIN as covariates.