GSM8076347: 5637SH-2.IP; Homo sapiens; OTHER
RNA extraction protocol：Total RNA was extracted using the TRIzol reagent (Invitrogen, USA). The RNA concentration of each sample was determined using a NanoDrop 2000 (Thermo Fisher, USA). The quality of the RNAs was subsequently determined by measuring the OD260/OD280 value, and RNA purity was confirmed by measuring the OD260/OD230 value. And the ratio of 260/280 between 1.8–2.0 and the ratio of 260/230 between 2.0–2.2 were considered to be pure RNA. Agarose gel electrophoresis was used for the integrity and quality testing of total RNA. Briefly, total RNA was used for removing the rRNAs with NEBNext® rRNA Depletion Kit (New England Biolabs, Inc., Massachusetts, USA) following the manufacturer's instructions. RNA libraries were constructed by using rRNA-depleted RNAs with TruSeq Stranded Total RNA Library Prep Kit (Illumina, USA) according to the manufacturer's instructions. And m1A RNA immunoprecipitation was performed with the GenSeqTM m1A-MeRIP Kit (GenSeq Inc., China) by following the manufacturer's instructions. Both the input samples without immunoprecipitation and the m1A IP samples were used for RNA-seq library generation with NEBNext® Ultra II Directional RNA Library Prep Kit (New England Biolabs, Inc., USA). The library quality was evaluated with BioAnalyzer 2100 system (Agilent Technologies, Inc., USA).