GSM7016398: miRNA-Seq cancer 1; Homo sapiens; ncRNA-Seq
Total RNA was extracted from endothelial cells  using RNeasy Mini Kit (Qiagen, Germany). The samples were freed from DNA using TURBO DNA-free kit (Thermo Fisher Scientific, USA) . RNA concentration and integrity was measured with UV-Vis spectrophotometry (Nanodrop  2000c,  Thermo  Fisher)  and  the  Fragment  Analyzer  System  using  the  DNF-471  RNA  Kit  (15  nt)  (Agilent,  Santa  Clara,  CA,  USA),  respectively.  As  per  manufacturer's  protocol,  the  libraries  were  generated using 100ng RNA samples with Small RNA-SeqLibrary Prep Kit (Lexogen,  Vienna, Austria,  #052UG128V0110). Then, cDNA library with adapters for sequencing was prepared and pooled in an  equimolar ratio. The concentration and the size distribution of the final lane mix was analyzed by Qubit  dsDNA HS assay (Thermo Fisher Scientific, Waltham, MA, USA) and by a Fragment Analyzer system  using the DNF-474 HS NGS Fragment Kit (1–6000 bp) (Agilent, Santa Clara, CA, USA). Dilution of  the lane mix (2 nm) was denatured and diluted to loading concentration for sequencing on a NextSeq  500  instrument  with  a  SR75  High  Output  Kit  (Illumina).  Differentially  expressed  miRNA  (DEmiRNAs) were classified according to −1 < logFC > 1, adjusted p-value—FDR (False Discovery  Rate) <0.05.