GSM9251739: Patient_15; Homo sapiens; RNA-Seq
16 patients peripheral blood samples were collected using EDTA as an anticoagulant. The samples were centrifuged at 1500 xg for 15 minutes, and the supernatant was subsequently collected. This supernatant underwent a second centrifugation at 2500 xg for 15 minutes to obtain Platelet-Poor Plasma (PPP). A volume of 350 μl PPP was mixed with 28 ml of phosphate-buffered saline (PBS) and then ultracentrifuged at 100000 xg for 1 hour at 4°C using a Beckman Optima XPN-100. The resulting pellet upon ultracentrifugation , containing extracellular content, was dissolved in 100 μl of PBS and then combined with 800 μl of TRIZOL LS Reagent (ThermoFisher). RNA was extracted through a phenol-chloroform method and cDNA libraries were prepared using the Smart-Seq V4 ultra-low input RNA kit from Clontech Laboratories, following the manufacturer's instructions. The amplified cDNA was further processed to create Illumina-compatible libraries according to the TruSeq Nano DNA sample preparation guide (Illumina). These libraries were then subjected to paired-end sequencing (2x75 cycles) on an Illumina HiSeq2500 platform.