GSM9092349: SV-HUC-1 cells, DCBQ treatment, RNA, rep1; Homo sapiens; RNA-Seq
The total RNA of each sample was extracted using the TRIzol reagent (Invitrogen, USA). The RNA was treated with RNase-free DNase I for 30 min. The poly(A) containing mRNA were purified using Oligo (dT) Beads from about 1 ug total RNA. The captured mRNA was firstly fragmented into 100-200 nt using divalent cations at elevated temperature. The fragmented mRNA was reverse-transcribed with SuperScript II and then converted to double-stranded cDNA using RNaseH and DNA Polymerase I by random priming. After purification, the double-stranded cDNA was subjected to blunt-ending, dA addition to 3'-end and adapter ligation.