GSM8172188: H69M cells, sheIF6, polysomal RNA, replicate 2; Homo sapiens; RNA-Seq
Polysome-bound RNA were extracted from sucrose based fraction after polysome profiling using TRIzol LS (ThermoFisher) according to manufacturer's protocol and were quantified by Nanodrop microvolume spectrophotometers (Thermo Fisher Scientific). Cytoplasmic RNA were extracted from cell lysate in hypotonic buffer (5 mM Tris-HCl pH 7.5, 2.5 mM MgCl2, 1.5 mM KCl, 100 μg.ml-1 cycloheximide, 2 mM DTT, 100 U/ml RNase inhibitor (Promega), 0.5% sodium deoxycholate, 0.5% Triton X-100) using TRIzol LS and were quantified by Nanodrop as well. Sequencing libraries were generated using NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, USA, Catalog #: E7530L) following manufacturer's recommendations