Illumina HiSeq 1000 sequencing; rRNA 2'-O-methylation profile of SNORD71 knockdown cells
rRNA 2'-O-methylation profile of SNORD71 knockdown cells
SNORD71 KD and CRISPR control SW1353 cells were cultured in DMEM/F-12 low glucose with GlutaMAX (Thermo Fisher Scientific) supplemented with 10% FCS (Sigma-Aldrich), 1% A-A (Thermo Fisher Scientific) at 37Â°C in a humidified atmosphere and 5% CO2.  Total RNA was isolated using the RNeasy Plus Mini Kit (Qiagen,74134) following the manufacturer's protocol. RNA quality was determined with a 2100 Bioanalyzer (Agilent) and RNA 6000 Nano Kit (Agilent, 5067-1511). All samples had RNA integrity (RIN) number greater than 9, which was considered sufficient for further analysis.  Total RNA was randomly fragmented by alkaline hydrolysis, RNA fragments were end-repaired and converted to library using NEBNext Small RNA Library kit.