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Single-molecule fluorescence in situ hybridization (FISH) of circular RNA CDR1as

Item Type:Article
Title:Single-molecule fluorescence in situ hybridization (FISH) of circular RNA CDR1as
Creators Name:Kocks, C., Boltengagen, A., Piwecka, M., Rybak-Wolf, A. and Rajewsky, N.
Abstract:Individual mRNA molecules can be imaged in fixed cells by hybridization with multiple, singly labeled oligonucleotide probes, followed by computational identification of fluorescent signals. This approach, called single-molecule RNA fluorescence in situ hybridization (smRNA FISH), allows subcellular localization and absolute quantification of RNA molecules in individual cells. Here, we describe a simple smRNA FISH protocol for two-color imaging of a circular RNA, CDR1as, simultaneously with an unrelated messenger RNA. The protocol can be adapted to circRNAs that coexist with overlapping, noncircular mRNA isoforms produced from the same genetic locus.
Keywords:circRNA Visualization, circRNA Quantification, FISH, Single-Molecule Detection, In Situ Hybridization, Absolute Quantification Of Gene Expression
Source:Methods in Molecular Biology
Series Name:Methods in Molecular Biology
Title of Book:Circular RNAs
Publisher:Springer / Humana Press
Page Range:77-96
Official Publication:https://doi.org/10.1007/978-1-4939-7562-4_7
PubMed:View item in PubMed

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