GSM4822807: Multiple myeloma cancer stem cells-U266 CSC DHA10_rep2; Homo sapiens; RNA-Seq
Total RNA from cells was extracted using TRIzol The cDNA library was constructed following the manufacturer's instructions of NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, E7530) and NEBNext Multiplex Oligos for Illumina (NEB, E7500). The enriched mRNA was fragmented into approximately 200nt RNA inserts, which were used to synthesize the first-strand cDNA and the second cDNA. The double-stranded cDNA were performed end-repair/dA-tail and adaptor ligation. The suitable fragments were isolated by Agencourt AMPure XP beads (Beckman Coulter, Inc.), and enriched by PCR amplification. At last, PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 and qPCR. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kitv3-cBot-HS(Illumia) according to the manufacturer's instructions. After cluster generation, the library preparations were sequenced on an Illumina platform at Biomarker Technologies(Beijing, China) and pair-end reads were generated.