Fig 1: Selective purification of ribosomal-depleted (RiboMinus) and ribosomal-enriched (RiboPlus) transcripts and sequencing overview.(A) Selective purification of distinct RNA populations was performed using the Ribominus Eukaryote kit for RNA-seq (#Ambion, A10837-08) according to the manufacturer’s ‘standard protocol’ instructions. Total RNA (containing rRNA and other RNA transcripts) is incubated with 5’ biotin label probes complementary to rRNA RNA transcripts (28 S, 18 S, 5.8 S, and 5 S). Probes hybridize to rRNA and are separated using magnetic bead separation. Non-rRNA is purified using ethanol precipitation, whereas bead-bound rRNA is purified using Trizol RNA isolation. (B) 1 µg of Total RNA (HEK293) and RiboPlus and 150 ng of Ribominus were loaded on 1% TBE agarose gel stained with ethidium bromide (in duplicates). Both total RNA and RiboPlus (rRNA-enriched) contained bands corresponding to 28 S, 18 S, and 5 S rRNA, whereas RiboMinus contained a band corresponding to lower fragment length. (C–F) The box plot represents total read generated (C), total mapped reads (D), total gene counts (E), and total transcript count (F) per condition (from two replicates), per time point.