title: GSM9251738: Patient_14; Homo sapiens; RNA-Seq
library_construction_protocol: 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.