The h/m/r riboPOOL for Ribo-Seq samples is a rRNA depletion tool for ribosome profiling samples. The h/m/r riboPOOL for Ribo-Seq samples was designed to evenly deplete the 30-mer rRNA contaminants, which occur during ribosome profiling. The h/m/r riboPOOL for Ribo-Seq samples doesn’t just cover the entire human/mouse and rat rRNA, it also takes the abundancy of different rRNA contaminants into consideration. Meaning, for extremely abundant rRNA contaminants the oligo ratio was adjusted. This thorough design approach leads to depletion efficiencies between 53%-75%. Moreover, no bias was introduced when tested on human ribosome profiling samples. Therefore, the h/m/r riboPOOL for Ribo-Seq samples is the world’s first efficient rRNA depletion solution for ribosome profiling.