GSM5292232: NHEM_rep4 (miRNA-seq); Homo sapiens; miRNA-Seq
Cells were trypsinized from a T75 cell culture flask and total RNA including small RNAs was isolated using the miRNeasy Mini Kit from Qiagen (Hilden, Germany) according to the manufacturer's manual. In short, an adenylated adapter was ligated to the 3' end of the total RNA by a truncated T4 RNA Ligase 2, followed by the ligation of a 5' RNA-adapter by T4 RNA Ligase 1 (NEB). After reverse-transcription using a specific primer, the cDNA was amplified by PCR, gel-fractionated and bands or regions corresponding to miRNA inserts were cut out. PCR products from these gel slices were eluted in elution buffer, precipitated and dissolved in water. The quality of the libraries was assessed by qPCR and Bioanalyzer measurements.