title: GSM9012225: HCT15NC_2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted using Trizol reagent kit (Invitrogen, Carlsbad, CA,USA) accordingto the manufacturer's protocol. RNA quality was assessed on an Agilent 2100 Bioanalyzer(Agilent Technologies, Palo Alto, CA, USA) and checked using RNase free agarose gelelectrophoresis. After total RNA was extracted, mRNA was enriched by Oligo(dT)beads.Then the enriched mRNA was fragmented into short fragments using fragmentation buffer and reverslytranscribed into cDNA by using NEBNext Ultra RNA Library Prep Kit for Illumina(NEB#7530,New England Biolabs, Ipswich, MA, USA).The purified double-stranded cDNA fragments were end repaired, A base added, and ligated to Illumina sequencing adapters.The ligation reaction was purified with the AMPure XP Beads(1.0X).Ligated fragments were subjected to size selection by agarose gel electrophoresis and polymerase chain reaction(PCR) amplified.The resulting cDNA library was sequenced using Illumina Novaseq6000 byGene Denovo Biotechnology Co. (Guangzhou, China).