GSM8586506: UBS_KO1_2; Homo sapiens; OTHER
Total RNA was extracted using TRIzol reagent (TIANGEN).Polyadenylated RNA was separated from total RNA using Oligo dT Magnetic Beads (Vazyme).Bisulfite treatment was performed using UBS-2.In brief, 45 µL of UBS-2 reagent was preheated to 98 °C for 5 min in a PCR instrument, and then approximately 100 ng of poly(A) RNA in 5 µL of water was added and mixed well. The mixture was incubated at 98 °C for 9 min with the lid temperature set to 105 °C. After cooling to room temperature, desulphonation was accomplished on the Zymo-Spin IC columns following the manufacturer's protocol The quantity of the converted RNA was determined using the Qubit. Libraries were then constructed using the Stranded mRNA-seq Lib Prep Module for Illumina (ABclonal).