GSM5267184: MDA231-Rab27a KD EV rep2; Homo sapiens; miRNA-Seq
TRIZOL reagent (Thermo Fisher Scientific) was used to extract total RNA from cells, whereas TRIZOL LS reagent (Thermo Fisher Scientific) was used to extract total RNA from EVs following the manufacturer's protocol. RNA pellet was dissolved in RNase-free water, and subjected to library construction and deep sequencing. Each sample was independently subjected to library preparation and deep sequencing.  All small RNAs of 15–52 nts were selected and sequenced using the Illumina system, following the manufacturer's protocol (Illumina Inc., San Diego, CA).  Library preparation, as well as cluster generation and deep sequencing, was performed according to the 5' ligation-dependent (5′ monophosphate-dependent) manufacturer's protocol (Digital Gene Expression for small RNA; Illumina).  Small RNAs were size-selected between 17 and 52 nt according to the single-stranded DNA marker in the small RNA sequencing kit (Illumina).  The library was quantified using picoGreen and qPCR.  Sequencing was performed on a Genome Analyzer IIx (Illumina), and image processing and base calling were conducted using Illumina's pipeline.