title: GSM3214182: RP_denr2_1; RPF-seq; Denr shRNA2, rep1; Mus musculus; RNA-Seq
library_construction_protocol: Cells were lyzed in 1 ml lysis buffer (150 mM NaCl, 20 mM Tris-HCl pH7.4, 5 mM MgCl2, 5 mM DTT, 100 μg ml-1 cycloheximide, 1% Triton X-100, 0.5% Sodium deoxycholate, complete EDTA-free protease inhibitors (Roche) ). Lysates were incubated for 10 min on ice and cleared by centrifugation at 1000 x g, 4°C for 3 min in a tabletop centrifuge. Supernatants were flash-frozen and stored in liquid nitrogen.  Total RNA-seq libraries were generated from the lysate pool. For ribosome profiling libraries, lysate pools were digested with 5 U ART-Seq nuclease per OD for 45 min at room temperature and passed over size exclusion columns for ribosome purification prior to RNA extraction and library preparation. ART-Seq (Illumina)
attributes.tag: GEO Accession
attributes.value: GSM3214182