GSM2577090: MDA-MB-468 CSC; Homo sapiens; miRNA-Seq
Total RNA was isolated from non-TNBC stem cells (BT474) and TNBC stem cells (MDA-MB-468) using Trizol reagent (Ambion, Carlsbad, CA), and then assessed by an Agilent 2100 BioAnalyzer (Agilent Technologies, Palo Alto, CA). Total RNA of each sample was used to prepare the miRNA sequencing library, which included the following steps: 1) 3'-adaptor ligation; 2) 5'-adaptor ligation; 3) cDNA synthesis; 4) PCR amplification; 5) size selection of ~135-155 bp PCR amplified fragments (corresponding to ~15-35 nt small RNAs). The libraries were denatured as single-stranded DNA molecules, captured on Illumina flow cells, amplified in situ as clusters and finally sequenced for 51 cycles on Illumina HiSeq per the manufacturer's instructions.