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Bio-protocol_Nadal_2019.pdf (5.83 MB)

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2019-09-05

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Si us plau utilitzeu sempre aquest identificador per citar o enllaçar aquest document: https://hdl.handle.net/2445/220565

Yeast Single-cell RNA-seq, Cell by Cell and Step by Step

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https://doi.org/10.21769/BioProtoc.3359

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Single-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.

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Transcripció genètica, Llevats

Matèries (anglès)

Genetic transcription, Yeast

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Articles publicats en revistes (Institut de Recerca Biomèdica (IRB Barcelona))

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NADAL RIBELLES, Mariona, ISLAM, Saiful, WEI, Wu, LATORRE DOMENECH, Pablo, NGUYEN, Michelle, NADAL CLANCHET, Eulàlia de, POSAS, Francesc, STEINMETZ, Lars m.. Yeast Single-cell RNA-seq, Cell by Cell and Step by Step. _Bio Protoc_. 2019. Vol. 9, núm. 17. [consulta: 23 de gener de 2026]. ISSN: 2331-8325. [Disponible a: https://hdl.handle.net/2445/220565]

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