GSM7663032: HCC1143-2; Homo sapiens; miRNA-Seq
Total RNA was isolated from cells using miRNeasy Mini Kit (QIAGEN). Briefly, cells grown at 80\% confluence in a 100 mm cell culture dish were washed with PBS and collected in a 1.5 ml centrifuge tube. Cells suspended in approximately 200ul of PBS were sonicated under RNase-free conditions. Then, the lysate was treated with 700 ul of QIAzol (QIAGEN) and 140 ul of chloroform to isolate nucleic acids in the aqueous phase. RNA was extracted following the manufacturer's instruction and eluted with 30 uL of nucleic acid-free water. Finally, one ul of RNase inhibitor was added to the solution. The RNA quality and concentration were measured by nanodrop (Thermo Fisher Scientific) and Agilent 2100 Bioanalyzer (Agilent). QIAseq miRNA Library Kits (QIAGEN) and QIAseq miRNA NGS 12 Index IL (QIAGEN) were used for library construction.