GSM5145406: Sample 27_Late AD-2; Homo sapiens; RNA-Seq
PBMCs were purified by Ficoll gradient separation, according to the instructions of the manufacturer (Ficoll–PaqueTM Plus, GE Healthcare, Sweden). In short, 3 mL of Ficoll-Paque gradient was pipetted into two 15-ml centrifuge tubes. The blood was diluted with a ratio 1:1 in phosphate-buffered saline (PBS) and carefully layered over the Ficoll-Paque gradient. The tubes were centrifuged for 30 min at 400 × g. The cell interface layer was harvested carefully (8 ml/tube), and the cells were washed twice in PBS (for 10 min at 100 × g). PBMCs were resuspended in 1 mL complete culture medium (DMEM medium supplemented with 10 % newborn calf serum, 1 % PS). RNA libraries were prepared for sequencing using standard Illumina protocols