GSM5234148: shFTO_Pre_Rep2; Mus musculus; RNA-Seq
Total RNA was extracted using TRIzol Reagent and MeRIP-seq samples were immunoprecipitated by m6A antibody. Samples were first dephosporylated and 40-60nt region was selected according to 15% polyacrylamide TBE-urea gel separation. Desired RNA species were excised and soaked overnight followed by ethanol precipitation. the purified RNA was processed for Poly-(A) tailing reaction and reverse transcription afterwards. Eventually, PCR amplification was proceeded using Phusion High-Fidelity DNA Polymerase.