title: GSM5511479: Sh-NC rep2; Homo sapiens; OTHER
library_construction_protocol: Size exclusion columns (illustra MicroSpin S-400 HR Columns; GE Healthcare; catalog no. 27-5140-01) were   equilibrated with 3 mL of polysome buffer by gravity flow and centrifuged at 600g for 4 min at room   temperature. 100μL of digested RFs were added to the column and centrifuged at 600g for 2 min. Next,   10μL 10% (wt/vol) SDS was added to the elution, and RFs with a size greater than 17nt was isolated   according to the RNA Clean and Concentrator-25 kit (Zymo Research;R1017) After obtaining ribosome footprints above, Ribo-seq libraries were constructed using NEBNext® Multiple   Small RNA Library Prep Set for Illumina® (catalog no.E7300S, E7300L). Briefly, adapters were added to both ends of RFs, followed by reverse transcription and PCR amplification. The 140-160bp size PCR products   were enriched to generate a cDNA library and sequenced using Illumina HiSeqTM 2500