title: GSM4255910: HepG2 cells with esculetin; Homo sapiens; RNA-Seq
library_construction_protocol: According to the manufacturer's instructions (Invitrogen), total RNA was extracted from HepG2 cells with or without TRIzol® Reagent, and genomic DNA was removed with using DNase I (TaKara). According to the TruSeqTM RNA sample preparation Kit of Illumina, we prepared the RNA-seq transcriptome library by using 5 μg of total RNA. For a short timeperiod of time, we isolated mRNA by oligo(dT) beads and RNA was fragmented through a fragment buffer. The SuperScript double-stranded cDNA synthesis kit was used to synthesize Double-stranded cDNA. Based on Illumina's library construction protocol, the synthetic cDNA was then terminal repaired, phosphorylated and added “'A”' base. The 200–300 bp cDNA target fragments size on 2% Low Range Ultra Agarose were selected, and 15 PCR cycles were amplified using Phusion DNA polymerase (NEB). Illumina HiSeq X Ten (2 × 150bp read length) was sequenced using the paired-end RNA-seq sequencing library after quantification by TBS380.
attributes.tag: GEO Accession
attributes.value: GSM4255910