title: GSM7065288: HCT15shNSUN2-1.IP; Homo sapiens; OTHER
library_construction_protocol: We extracted total RNA using Trizol reagent (Invitrogen, California, USA) following the manufacturer's instructions. Use the Ribo-Zero rRNA Removal Kit (Illumina, Inc., CA, USA) to reduce rRNA content. The quality of RNA is assessed using its OD260/OD280 ratio, which is a measure of nucleotide-to-protein ratio based on optical density measured using spectrophotometry. RNA purity with OD260/OD280 values in the range of 1.8-2.1 is considered acceptable, and RNA extracted from all samples meets this standard. Total RNA was cleaved into 100 base pair fragments using the GenSeqTM m5C RNA IP Kit (GenSeq Inc, China), followed by m5C immunoprecipitation using the NEBNext® Ultra II Directed RNA Library Preparation Kit (New England Biolaboratories, USA), followed by RNA-seq library generation.The cDNA library was evaluated using the Bioanalyzer 2100 System (Agilent Technologies USA) and the library was sequenced using an Illumina Hiseq instrument with 150 bp pairing readings.