RIP-seq of melanoma stem cells
Briefly, the rRNAs were removed by ribo-zerotm kit (Epicentre, France). Subsequently fragmentation buffer was added to break the RNA into short segments of 250-300bp, followed by the synthesis of cDNAs with random hexamers. After purification with AMPure XP beads, the double-stranded cDNAs were added with A tails and the connection of sequencing joints. The cDNA library was enriched by PCR. Sequencing was performed. After assembly of RNA-seq data, the raw data was processed to remove the sequences of adapter, ploy-N and low-quality reads.