Optimized protocol for single-molecule RNA FISH to visualize gene expression in S. cerevisiae

Single-molecule RNA fluorescence in situ hybridization (smFISH) allows subcellular visualization, localization, and quantification of endogenous RNA molecules in fixed cells. The spatial and intensity information of each RNA can be used to distinguish mature from nascent transcripts inside each cell...

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Detalles Bibliográficos
Autores principales: Patel, Heta P., Brouwer, Ineke, Lenstra, Tineke L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8264745/
https://www.ncbi.nlm.nih.gov/pubmed/34278333
http://dx.doi.org/10.1016/j.xpro.2021.100647
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author Patel, Heta P.
Brouwer, Ineke
Lenstra, Tineke L.
author_facet Patel, Heta P.
Brouwer, Ineke
Lenstra, Tineke L.
author_sort Patel, Heta P.
collection PubMed
description Single-molecule RNA fluorescence in situ hybridization (smFISH) allows subcellular visualization, localization, and quantification of endogenous RNA molecules in fixed cells. The spatial and intensity information of each RNA can be used to distinguish mature from nascent transcripts inside each cell, revealing both past and instantaneous transcriptional activity. Here, we describe an optimized protocol for smFISH in Saccharomyces cerevisiae with optimized lyticase digestion time and hybrization steps for more homogenous results. For complete details on the use and execution of this protocol, please refer to Donovan et al. (2019).
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spelling pubmed-82647452021-07-16 Optimized protocol for single-molecule RNA FISH to visualize gene expression in S. cerevisiae Patel, Heta P. Brouwer, Ineke Lenstra, Tineke L. STAR Protoc Protocol Single-molecule RNA fluorescence in situ hybridization (smFISH) allows subcellular visualization, localization, and quantification of endogenous RNA molecules in fixed cells. The spatial and intensity information of each RNA can be used to distinguish mature from nascent transcripts inside each cell, revealing both past and instantaneous transcriptional activity. Here, we describe an optimized protocol for smFISH in Saccharomyces cerevisiae with optimized lyticase digestion time and hybrization steps for more homogenous results. For complete details on the use and execution of this protocol, please refer to Donovan et al. (2019). Elsevier 2021-07-06 /pmc/articles/PMC8264745/ /pubmed/34278333 http://dx.doi.org/10.1016/j.xpro.2021.100647 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Patel, Heta P.
Brouwer, Ineke
Lenstra, Tineke L.
Optimized protocol for single-molecule RNA FISH to visualize gene expression in S. cerevisiae
title Optimized protocol for single-molecule RNA FISH to visualize gene expression in S. cerevisiae
title_full Optimized protocol for single-molecule RNA FISH to visualize gene expression in S. cerevisiae
title_fullStr Optimized protocol for single-molecule RNA FISH to visualize gene expression in S. cerevisiae
title_full_unstemmed Optimized protocol for single-molecule RNA FISH to visualize gene expression in S. cerevisiae
title_short Optimized protocol for single-molecule RNA FISH to visualize gene expression in S. cerevisiae
title_sort optimized protocol for single-molecule rna fish to visualize gene expression in s. cerevisiae
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8264745/
https://www.ncbi.nlm.nih.gov/pubmed/34278333
http://dx.doi.org/10.1016/j.xpro.2021.100647
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