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| Item Type: | Article |
|---|---|
| Title: | Single-molecule fluorescence in situ hybridization (FISH) of circular RNA CDR1as |
| Creators: |
Kocks, C.  ORCID: https://orcid.org/0000-0003-1749-3334, Boltengagen, A. ORCID: https://orcid.org/0000-0002-8356-9766, Piwecka, M. ORCID: https://orcid.org/0000-0003-4104-6577, Rybak-Wolf, A. ORCID: https://orcid.org/0000-0002-1192-5290 and Rajewsky, N. ORCID: https://orcid.org/0000-0002-4785-4332
|
| 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 |
| ISSN: | 1064-3745 |
| ISBN: | 978-1-4939-7561-7 |
| Publisher: | Springer / Humana Press |
| Volume: | 1724 |
| Page Range: | 77-96 |
| Date: | 2018 |
| Official Publication: | https://doi.org/10.1007/978-1-4939-7562-4_7 |
| PubMed: | View item in PubMed |
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