DM3
The isolated footprinted RNA was depleted of rRNA using Illumina Ribo-Zero Gold kit (Illumina). The RNA was separated on polyacrylamide gel to purify ribosome-protected fragments (RPF). Fragments (27-30 nt) of RNA were eluted from the gel and used for library construction with Truseq small RNA library kit (RS-200-0012 Illumina). Ribo-seq was sequenced on an Illumina novaseq with paired-end 150 bp reads.