GSM8560360: rnaseq_control_rep1; Homo sapiens; RNA-Seq
For RNA-seq cells were harvested with Trizol followed by chloroform-based RNA exraction. For Ribosome profiling- Cells were treated with CHX 100ug/mL for 1min, followed by PBS wash supllemented with 100ug/mLCHX and lysed with lysis buffer (12.5 mM Tris, pH 7.0; 7.5mM Tris, pH 8.0; 150 mM NaCl; 5 mM MgCl2; 100 μg/mL cycloheximide; 1 mM DTT; 1% (v/v) Triton X-100; 20 U/mL DNase). cell lysate were collected by centrifugation 10min 15,000xg 4c. Lysate were treated 45min with 250U RNAse I, and quenched with SUPERase Inhibitor, and ribosome pellet was collected by 1h 100k rpm centrifugation  at 4c with TRIZOL, and chloroform-based separation.