title: GSM8264824: SITW019_1; Mus musculus; OTHER
library_construction_protocol: Ribo-Calibration: Cells in a 10-cm dish were lysed with 600 µl of lysis buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM MgCl2, 1% Triton X-100, 1 mM DTT, 100 µg/ml cycloheximide, and 100 µg/ml chloramphenicol) and treated with 25 U/ml TURBO DNase (Thermo Fisher Scientific). Then, the lysate was clarified by centrifugation at 20,000 × g for 10 min at 4°C. Ribo-Calibration: The spike-in mixture of Rluc-disome and Fluc-trisome were added to the lysate. For Ribo-Seq, RNase treatment, ribosome pelleting, Ribo-FilterOut, size selection, ligation of linker with the antisense sequence of T7 promoter, rRNA depletion, in vitro transcription, ligation of second linker, reverse transcription, and PCR amplification. For RNA-Seq, the libraries were prepared by a SEQuoia Express Standard RNA Library Prep Kit (Bio-Rad).