RNA-sequencing (RNA-Seq) protocols and bioinformatic pipelines are designed to streamline downstream analyses on sequences believed to be the most important. Here, we have challenged this dogma by preserving ribosomal RNA (rRNA) in our samples and by lowering the minimal RNA size window of our small RNA-Seq analyses to 8 nt Overall design: Analyzing the content of the RNA size window between 8 and 30 nt of 13 different biological samples from 6 different organisms by small RNA-Seq.