| Chapter title |
Single-Molecule Fluorescence In Situ Hybridization (FISH) of Circular RNA CDR1as
|
|---|---|
| Chapter number | 7 |
| Book title |
Circular RNAs
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7562-4_7 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7561-7, 978-1-4939-7562-4
|
| Authors |
Christine Kocks, Anastasiya Boltengagen, Monika Piwecka, Agnieszka Rybak-Wolf, Nikolaus Rajewsky |
| 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. |
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