RNA molecules play important roles in virtually every cellular process. These functions are often mediated through the adoption of specific structures that enable RNAs to interact with other molecules. Thus, determining the secondary structures of RNAs is central to understanding their function and evolution. In recent years several sequencing-based approaches have been developed that allow probing structural features of thousands of RNA molecules present in a sample. Here, we describe nextPARS, a novel Illumina-based implementation of in vitro parallel probing of RNA structures. Our approach achieves comparable accuracy to previous implementations, while enabling higher throughput and sample multiplexing.

This work was supported by the Spanish Ministry of Economy and Competitiveness grants “Centro de Excelencia Severo Ochoa 2013–2017” (SEV-2012-0208); the European Regional Development Fund (ERDF) from the European Union and European Research Council (ERC) Seventh Framework Programme (FP7/2007–2013) (ERC-2012-StG-310325); and the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement (H2020-MSCA-ITN-2014-642095).