GSM5850941: Monte024; Homo sapiens; RNA-Seq
A single sample of blood (2.5-5ml) was collected for peripheral blood transcriptome analysis after consent in a PAXgene tube with RNA stabilization reagents. The date of sample collection was considered enrollment date. PAXgene samples from all patients were transported to the central laboratory at Mount Sinai and stored at -80C. RNA extraction and processing were done in batches of ten by 2 experienced operators. Total RNA was stored at -80C till sequencing. QIAamp RNA Blood Kit was used to extract high quality RNA from total blood (<1% DNA contamination; Qiagen, CA, USA). Total RNA (>500 ng per sample; 260/280 ratio = 2.07  0.08) was poly-A selected and amplified for library preparation and subsequent sequencing (Genewiz Inc, NJ, USA). 150bp paired-end sequencing was performed on the prepared libraries (Illumina, CA, USA) to generate 20-30 million reads per sample, and raw reads data was stored in FASTQ files for analysis.