Ribo-Seq, also referred to as ribosome profiling or ribosome footprinting, is a next-generation sequencing (NGS)-based transcriptomic research technique that measures translation comprehensively and quantitatively. Ribo-Seq detected the precise positions of ribosomes to determine which mRNAs are being actively translated, which allows for the profiling of the active ribosomes in a cell at a particular moment, as well as the location of the translation initiation sites, the complement of translated ORFs, the specific locations of ribosomes on the mRNA, and the translation rate of ribosomes.