GSM8245199: hAoSMCs, miR-15a mimic, rep1; Homo sapiens; RNA-Seq
RNA was harvested by pooling two wells of each condition to assure adequate RNA concentration. Extraction was performed using miRNeasy Tissue/Cells Advanced Mini Kit (Qiagen), following the manufacturer's protocol, including an on-column DNAse digestion step. mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using dUTP. The directional library was complete after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and Bioanalyzer for size distribution detection. The clustering of the index-coded samples was performed according to the manufacturer's instructions.