GSM6592462: DOK cell IP replicate 2; Homo sapiens; RIP-Seq
total RNA was isolated using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and quantified. RNA integrity was evaluated by Bioanalyzer 2100 (Agilent, CA, USA). Poly (A) RNA was purified using Dynabeads Oligo (dT)25-61005 (Thermo Fisher, CA, USA) and fragmentedinto about 100nt using Magnesium RNA Fragmentation Module (NEB, USA). The RNA fragments weredivided into two portions, one was kept as input and the otherwas used to enrich m6A-methylated RNA fragments by immunoprecipitation with m6A-specific antibody (Synaptic Systems, Germany). RNA-seq library preparation included double stranded cDNA synthesis, addition of A-tailing, adapter ligation, amplification of ligated products, and library purification. The paired-end sequencing (PE150) of libraries was performed on Illumina Novaseq™ 6000 platform