title: GSM9269783: HEK293T, Torin1-treated, Ribo-Seq, rep2; Homo sapiens; OTHER
library_construction_protocol: Cells were chilled on ice and washed with PBS supplemented with cycloheximide (100 µg/ml), lysed with buffer containing 20 mM Tris-HCl (pH 7.4), 150 mM NaCl, 5 mM MgCl2, 1 mM DTT, 1% Triton X-100, 100 µg/ml cycloheximide, and 25 U/ml TURBO DNase (Ambion). The lysate was divided into two parts for Ribo-Seq and RNA-Seq library preparation. Ribo-Seq libraries were obtained according to (McGlincy 2017, PMID: 28579404) using CircLigase II (Lucigen), custom adapter with UMI-sequence (5 random nucleotides  for deduplication), and NEBNext Multiplex Oligos for Illumina Index Primers Set (NEB). The rRNA depletion was performed using a RiboMinus™ Eukaryote Kit v2 (Thermo Fisher Scientific) according to the manufacturer's recommendations.