GSM5234110: Pro_rep1_rna; Homo sapiens; RNA-Seq
Three independent replicas of proliferating or senescent IMR90 were grown, harvested in ice-cold polysome isolation buffer supplemented with cyclohexamide. Approx. 15% of each lysate was kept for isolation of RNA and used for RNA-seq. Cell lysates digested with RNase1, ribosomes purified, RNA extracted, ribosome protected fragments (28-32nt) isolated by urea-PAGE then NGS libraries prepared by 3' linker ligation followed by reverse transcriptase with a circularisation primer (same as doi.org/10.1016/j.ymeth.2017.05.028).