Illumina HiSeq 1000 sequencing; miRNA expression profile in platelets after ex vivo activation
miRNA expression profile in platelets after ex vivo activation
Blood has been processed within 15 minutes from sampling and platelets isolated by differential centrifugation. Platelets were activated with ADP (20uM), Collagen (60ug/mL) or Thrombin Receptor Activating Peptide (TRAP, 25uM) for 120 minutes, under continuous stirring. Total RNA was extracted using RNA was extracted using mirVana Paris kit (life technologies) according to the manufacturer instructions. 1ug of total RNA was used in a library preparation according to the Illumina TruSeq small RNA sample preparation protocol (Illumina, USA). Libraries were multiplexed, by using Illumina indices set A, with indices 36 (RPI36), 38 (RPI38), 39 (RPI39) and 40 (RPI40) for T0 (control), ADP, TRAP and COLL treatment respectively.
Experimental Factor: compound
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