GSM6632448: EV_WI-38_IR_3; Homo sapiens; RNA-Seq
EVs were extracted by ultracentrifugation and total RNA was isolated from IR-treated cells and EVs with RNeasy Mini Kit and exoRNeasy Serum/Plasma Maxi kits (QIAGEN).  RNA-Seq libraries were prepared using the KAPA RNA Hyper Prep kit with RiboErase (HMR) to yield 200-300 bp insert size (Roche, Wilmington, MA).  The fragmented cDNA was ligated with adapters (Integrated DNA Technologies, Inc.), and amplified with KAPA HiFi HotStart ReadyMix (2X) and primers (10X) for 17 cycles.  Clean-up after amplification was performed with 1× KAPA Pure Beads.  Finally, the quality and quantity of the cDNA library were analyzed by Agilent Tapestation 4200 and Qubit 3.0 Fluorometer (Thermo Fisher Scientific Inc, Waltham, MA). The final libraries were subjected to 150 bp PE (paired-end) sequencing on an Illumina HiSeq sequencer (Illumina Inc., San Diego, CA).