title: GSM6670893: CD45Cells: Leucocytes; Homo sapiens; miRNA-Seq
library_construction_protocol: Peripheral blood samples from two healthy were collected into sterilize vacutainer blood collection tubes with K3-EDTA, pooled and processed within 6h after blood collection. Erythrocytes, leukocyte-platelet suspension and plasma fractions were collected after the first centrifugation of the whole blood sample at 1500 g for 10 minutes.  Platelets and leukocytes (CD45+cells) were obtained by highly sensitive fluorescence activated cell sorting. Subpopulations of plasma EVs were obtained using high-sensitivity fluorescence activated vesicle sorting. Additionally, 500 µl plasma sample collected after the whole blood centrifugation step described above was centrifuged at 16000 g for one hour at +4C, then 450 µl of the upper fraction supernatant (S16) was collected. Whereas, the pellet (P16) was resuspended in 100 µl PBS рН 7.4. Small RNA libraries were generated using Small RNA-SeqKit (Lexogen, Vienna, Austria) according to manufacturer's recommendations. The number of amplification cycles was 22 for sorted CD45+ cells and extracellular vesicle samples and 16 for other samples.