GSM5663144: H_Exp: HCC cell line [HBx overexpression]; Homo sapiens; ncRNA-Seq
A total of two samples from parental HepG2-H5298 and HepG2-H6679 cells were used for RNA extraction, performed as above (Trizol reagents, Invitrogen). The RNA amount and purity of each sample were quantified using NanoDropND-1000 (NanoDrop, Wilmington, DE, USA). The RNA integrity was assessed using Agilent 2100 with RIN number >7.0. Approximately 5 µg of total RNA was used to deplete ribosomal RNA according to the manuscript of Ribo-Zero™ rRNA Removal Kit (Illumina, San Diego, CA, USA). After removing ribosomal RNAs, the left RNAs were fragmented into small pieces using divalent cations under high temperatures. After digestion, reverse transcription, and amplification with PCR, cDNA was obtained, and the average insert size for the final cDNA library was 300 bp (±50 bp). At last, we performed a paired-end sequencing on an IlluminaHiseq 4000 (Illumina) following the vendor's recommended protocol.