GSM9138087: Control 1; Homo sapiens; miRNA-Seq
Total RNA, including miRNA, was extracted from plasma-derived EVs using the Total Exosome RNA & Protein Isolation Kit (Invitrogen, USA) employing acid-phenol:chloroform for organic extraction according to the manufacturer's protocol. Briefly, ethanol was added to the aqueous phase obtained from acid-phenol:chloroform extraction and passed through a glass-fiber filter cartridge, which immobilized the RNA. The filter was subsequently washed, and RNA, including the small RNA fraction, was eluted using a low ionic-strength buffer. RNA sequencing libraries were prepared using the KAPA RNA HyperPrep Kit with RiboErase (HMR) (Roche, Switzerland) following the manufacturer's protocol. Li-braries were sequenced on the Illumina NextSeq 500/550 platform using Mid Output kits (v2.5). Image processing, base calling, and demultiplexing were performed with NextSeq Control Software (Illumina, San Diego, CA, USA).