title: NextSeq 500 sequencing; RNA-Seq of effects of exercise-induced Extracellular Vesicles on an F344 rat (Rattus norvegicus) model of forced running wheel exercise
design: RNA-Seq of effects of exercise-induced Extracellular Vesicles on an F344 rat (Rattus norvegicus) model of forced running wheel exercise
library_construction_protocol: After 4 weeks of regular exercise, rats underwent brief anesthesia and of blood from the tail vein was taken and immediately put into K3 EDTA tubes and centrifuged for 15 min at 2000 x g at 4 degrees C. This step was repeated for the same animals immediately after 1h of running. On the same day, a blood sample was collected from the sedentary rats following the same protocol.  Plasma was collected and immediately frozen for further EV isolation. RNA was extracted from plasma EVs using miRNeasy micro kit (Qiagen, USA) according to the manufacturer's protocol. The concentration and quality of the obtained RNA were analyzed on Agilent Bioanalyzer with Agilent RNA 6000 Pico chip (Agilent Technologies, Germany). RNA libraries were constructed using CleanTag® Small RNA Library Prep Kit (Trilink Biotechnologies, USA), the quality and concentration of obtained libraries were analyzed on Agilent Bioanalyzer using Agilent High Sensitivity DNA chip (Agilent Technologies, Germany). The libraries were cleaned using Blue Pippin DNA Size Selection with 3% gel Blue Pippin Cassette (Sage Science, USA) setting tight target length to 140 bp thus selecting fragments with size in tight range to 140 bp (133–147 bp).