GSM5610294: TSC control replicate 1; Macaca mulatta; RNA-Seq
Each EV sample was diluted in 5 volumes Qiazol and incubated for 5 mins at RT prior to freezing at -80°C until RNA extraction. The miRNeasy serum/plasma kit (Qiagen, Cat #217184) was used to isolate total RNA from EVs following the manufacturer's protocol with minor adaptations. The Qiazol with Chloroform was overlaid onto phase maker tubes (ThermoFisher, Cat #A33248) and then spun at 4°C at 16,000 x g for 15 mins. In addition, two RPE washes of 500 µl were applied prior to RNA elution. All samples were eluted in 30 µl RNAse free water and the initial elution was placed back on the column and reeluted to increase RNA concentration. Sample concentration and purity was determined with the NanoDrop One (ThermoFisher, Cat # ND-ONE-W). The poly(A) RNA cDNA sequencing library was prepared following Illumina's TruSeq-stranded-mRNA sample preparation protocol.