title: GSM8761946: Stiff EV mRNA biol rep 2; Homo sapiens; RNA-Seq
library_construction_protocol: EVs were isolated from culture media samples using an insulator-based dielectrophoresis approach. Total RNA was extracted from purified EVs using the miRNeasy Micro kit as per the manufacturer's suggested protocol. Size distribution and estimated RNA concentration were measured using the Agilent 6000 Pico Kit using Bioanalyzer. Directional whole exosome RNA-seq was performed by the Genomics, Epigenomics and Sequencing Core (GESC) at the University of Cincinnati. To summarize, the quality of total RNA was quality control analyzed by Bioanalyzer. A total of ~10 ng RNA was input for library preparation using NEBNext Ultra II Directional RNA Library Prep kit under PCR cycle number 9. Library quality control and quantification were performed via Qubit quantification, and individually indexed libraries were proportionally pooled and sequenced using NextSeq 2000 Sequencer under the PE 2x61 bp setting to generate about 44M reads. Upon sequencing completion, fastq files were generated via Illumina BaseSpace Sequence Hub for downstream data analysis.