title: GSM5050950: CTL 3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from at least three biological replicates of controls and GDF11 treated (100ng/ml, 24 h) HepG2 cells with TRIzol Reagent (Invitrogen, CA, US) and column separation using a DNeasy Blood & Tissue Kit (Qiagen, Germany), according to the manufacturer's instructions. RNA integrity was assessed using Agilent RNA 6000 Nano Kit and Agilent 2100 Bioanalyzer (both Agilent Technologies, CA, US). Samples with RIN≥8.5 were further processed for sequencing. Indexed libraries were prepared from 250ng purified RNA using a NEB Next Ultra II Directional RNA Library Prep Kit (Illumina, UK), according to the manufacturer's instructions. The libraries were pooled in equimolar amounts(with a final concentration of 2 nmol/L), subjected to cluster generation and sequenced on an Illumina MiSeq System (Illumina) in a 1x75 format. Quality control was performed by FastQC and MultiQC. Short reads related to the sequencing of control and GDF11 treated HepG2 cells were aligned against the hg19 genome assembly, using STAR (ver. 2.6) with standard parameters.