Protocol for single-nucleus RNA sequencing of adult C. elegans neurons

Single-nucleus sequencing, specifically, remains the gold standard for characterizing difficult-to-dissociate cell types, such as neurons in complex brain tissue, both in mammals and in Caenorhabditis elegans . Here, we present a protocol for single-nucleus RNA sequencing of adult C. elegans neurons. This protocol describes the steps for creating a high-quality single-nucleus suspension from C. elegans hermaphrodite and male neurons, the nuclei sorting conditions, RNA sequencing library preparation, and analysis pipelines. For complete details on the use and execution of this protocol, please refer to St. Ange et al. ¹ and Morillo et al.