Abstract
The current versions of the microRNA databases MiRgeneDB, miRBase, and PmiREN contain annotations for a total of 358 different species. Public repositories, however, host small RNA sequencing data for over 800 species. This discrepancy implies that microRNA research is also very active in species that neither have an available high-quality genome assembly nor annotations for microRNAs or other types of noncoding genes. These cases are particularly challenging to analyze because reference sequences need to be collected from different sources and processed and formatted appropriately so that the dedicated small RNA analysis tools can make use of them. In this protocol we describe how small RNA sequencing data can be easily analyzed by means of a dockerized version of the well-established sRNAtoolbox/sRNAbench small RNA tools. We outline the analysis of two publicly available datasets to demonstrate basic aspects like the preparation of the local database, expression profiling, or differential expression analysis as well as more advanced features such as quantification of exogenous RNA content and data analysis in non-model species.
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