Nadal Ribelles, MarionaIslam, SaifulWei, WuLatorre Domenech, PabloNguyen, MichelleNadal Clanchet, Eulàlia dePosas, FrancescSteinmetz, Lars M.2025-04-242025-04-242019-09-052331-8325https://hdl.handle.net/2445/220565Single-cell RNA-seq (scRNA-seq) has become an established method for uncovering the intrinsic complexity within populations. Even within seemingly homogenous populations of isogenic yeast cells, there is a high degree of heterogeneity that originates from a compact and pervasively transcribed genome. Research with microorganisms such as yeast represents a major challenge for single-cell transcriptomics, due to their small size, rigid cell wall, and low RNA content per cell. Because of these technical challenges, yeast-specific scRNA-seq methodologies have recently started to appear, each one of them relying on different cell-isolation and library-preparation methods. Consequently, each approach harbors unique strengths and weaknesses that need to be considered. We have recently developed a yeast single-cell RNA-seq protocol (yscRNA-seq), which is inexpensive, high-throughput and easy-to-implement, tailored to the unique needs of yeast. yscRNA-seq provides a unique platform that combines single-cell phenotyping via index sorting with the incorporation of unique molecule identifiers on transcripts that allows to digitally count the number of molecules in a strand- and isoform-specific manner. Here, we provide a detailed, step-by-step description of the experimental and computational steps of yscRNA-seq protocol. This protocol will ease the implementation of yscRNA-seq in other laboratories and provide guidelines for the development of novel technologies.22 p.application/pdfeng(c) Nadal Ribelles, Mariona et al., 2019Transcripció genèticaLlevatsGenetic transcriptionYeastinfo:eu-repo/semantics/article2025-04-22info:eu-repo/semantics/openAccess623273033654857