GSM6701569: MGG4_GSC_RiboMeth; Homo sapiens; OTHER
Cell culture medium were removed and RNA was harvested using Trizol reagent. For RiboMeth-seq, total RNA was fragmented in 50 mM sodium carbonate/bicarbonate buffer (pH 9.2) at 95°C. Next, Fragmented RNA was 3′-end dephosphorylated and purified with the RNA Clean & Concentrator-5 kit (Zymo Research). RNA was then 5′-end phosphorylated with T4 polynucleotide kinase (NEB) and purified again. For uvRIP-seq, RNA were harvested with Trizol reagent after NOP58 UV-crosslinked immunoprecipitaion. For RiboMeth-seq, sequencing libraries were prepared with the VAHTSTM Small RNA Library Prep Kit for Illumina® (Vazyme) following the manufacturer's instruction. For RNA-seq, RNA libraries were prepared for sequencing using VAHTSTM Total RNA-seq (H/M/R) Library Prep Kit for Illumina (Vazyme) with total RNA. Ribo-seq: ​Ribosome profiling was performed using Ribosome Profiling Kit.  RPFs  was extracted using RNA clean&ConcentratorTM-5 kit.     EpiTM RiboRNA Depletion Kit  was used for rRNA depletion.  Sequencing libraries were constructed using QIAseq miRNA Library kit.