title: GSM6723864: SCC61 cells, rep1; Homo sapiens; OTHER
library_construction_protocol: The total RNA from 2 μg of the lysate was purified with the RNA Clean and Concentrator-5 kit (Zymo Research). 15 μg of lysate was digested with RNase I (Thermofisher) at 21°C for 45 min. The digested lysate was purified on a MicroSpin S-400 HR Sephacryl column (GE Healthcare). Ribosome-protected fragments (RPFs) of 28-30 nt were selected by loading the footprints on a Novex TBE-Urea gel (15%) (Thermofisher) and performing gel electrophoresis. The gel parts corresponding to the fragments of interest were excised and let to diffuse in molecular grade water supplemented with ammonium acetate and SDS overnight at 4°C. The Small RNA Library prep kit (Lexogen) was used to generate the library. Ribo-seq; High coverage sequencing was performed on the Illumina NovaSeq6000, SE50 single index configuration.