RNA-seq of homo sapien: adult blood malignant PBMCs: Sample S15
Peripheral blood mononuclear cells (PBMCs) were isolated from blood of patients and healthy controls by Ficoll-Hypaque gradient centrifugation. Genomic DNA was extracted utilizing QIAamp Micro Kit (Qiagen). RNA was extracted using RNeasy Plus Micro Kit (Qiagen) from non-stimulated malignant cells and stimulated with anti-CD3/CD28 beads (1:1 ratio, Dynabeads human T-activator CD3/CD28, Life Technologies). FFPE samples were obtained by needle cores taken from tumor rich areas of each block. DNA and RNA were extracted using QIAamp FFPE DNA (Qiagen) or FFPE RNA/DNA Purification Plus (Norgen) kits respectively. For frozen tumor samples, tumor biopsies were obtained and stored immediately in RNAlater (ThermoFisher) overnight at 4°C. The following day, samples were transferred to -80°C until processing. DNA and RNA were extracted using AllPrep DNA/RNA Mini (Qiagen).