RNA-seq of homo sapiens : melanoma cell line
Messenger RNA was purified from the total RNA using poly T oligo-attached magnetic beads. After fragmentation, the first-strand cDNA was synthesized using random hexamer primers. Next, second-strand cDNA was synthesized using dUTP. The directional library was prepared 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 a bioanalyzer for size distribution detection.