title: GSM5105027: Disomes – Wild type; Severe acute respiratory syndrome coronavirus 2; OTHER
library_construction_protocol: 50µl Rabit reticulocyte lysate were digested in 1 volume of MNase buffer (5 mM CaCl2, 100 mM NaCl, 20 mM Tris-HCl pH 7.5, 10 mM MgCl2, 5 mM DTT) with 750 U MNase (NEB) for 45 min at room temperature. The samples were further digested in 2 volumes of lysis buffer (150 mM NaCl, 20 mM Tris-HCl pH 7.4, 5 mM MgCl2, 5 mM DTT, 1% Triton X-100, 0.5% Sodium deoxycholate, complete EDTA-free protease (Roche) and 40 U/ml RNasin plus (Promega))  with 100 U RNase I (Ambion) for 45 min at room temperature. Digested samples were passed over size exclusion columns for ribosome purification prior to RNA extraction and library preparation. Sequencing libraries were generated using custom-designed riboPOOL (SiTools) and protocol adapted from previously reported protocols (Arpat et al., 2020; PMID 32703885 and McGlincy and Ingolia, 2017; PMID 28579404).