GSM7962078
The cultures were harvested by rapid filtration and freezing in liquid nitrogen. Cells were lysed with 1 M NaCl-containing lysis buffer and 25 AU of extracted RNA lysates was pelleted over a 1-ml sucrose cushion (20 mM Tris [pH 7.5], 500 mM NH4Cl, 0.5 mM EDTA, 1.1 M sucrose) using a TLA 100.3 rotor at 65,000 rpm for 2 h. Pellets were resuspended in 200 ml of lysis buffer and the RNA was digested with MNase. We followed the described protocol to construct libraries (Mohammad F, Green R, Buskirk AR: A systematically-revised ribosome profiling method for bacteria reveals pauses at single-codon resolution. Elife 2019, 8.) Ribo-Seq