9CSC28_shNC_ribo seq_CUHKIDD_A5
The Ribo-sequencing was followed as described in an established strategy. Briefly, Cycloheximide (Sigma, St Louis, MO, USA) was added to cell cultures medium to a final concentration of 100 g/ml. To prepare ribosome footprints (RFs), 10L of RNase I (NEB, Ipswich, MA, USA) and DNase I (NEB, Ipswich, MA, USA) were added to lysate. RFs were purified using magnet beads (Vazyme, Nanjing, Jiangsu, China). After obtaining the RFs above, Ribo-seq libraries were constructed by NEBNext Multiple Small RNA Library Prep Set for Illumina (catalog no. E7300S, E7300L). The 140-160bp size PCR products were enriched to generate a cDNA library and sequenced using Illumina HiSeqTM X10.