GSM8305467: shFTO-2_rep2; Rattus norvegicus; RNA-Seq
Total RNA was isolated and purified using TRIzol reagent (Invitrogen, USA) following the manufacturer's procedure. Cleaved RNA fragments were reverse-transcribed to create the cDNA by SuperScript II Reverse Transcriptase (Invitrogen, cat. 1896649, USA), which were next used to synthesise U-labeled second-stranded DNAs with E. coli DNA polymerase I (NEB, cat.m0209, USA), RNase H (NEB, cat.m0297, USA) and dUTP Solution (Thermo Fisher, cat.R0133, USA). At last, we performed sequencing using Illumina NovaSeq6000 PE150 (LC-Bio Technology Co., Ltd., Hangzhou, China).