GSM8765400: HAP1 cells, 5-azaC treated, biol rep1; Homo sapiens; OTHER
Cells were lysed with a buffer containing TritonX-100 and cycloheximide to arrest translation. Lysates were treated with RNase I. Ribosomes were then pelleted through a sucrose cushion.  and RNA was isolated by phenol extraction. Ribosome-protected mRNA footprints of 26 - 32 nt were isolated by gel size selection. Library construction was carried out essentially as described (McGlincy & Ingolia, 2017) with minor modifications. rRNA was depleted with rRNA-tRNA (RiboSeq) riboPOOLs (siTOOLs) following the 3' adapter ligation step.