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Genome-Wide Study of mRNA Isoform Half-Lives
- Source :
- Methods in Molecular Biology ISBN: 9781493930661
- Publication Year :
- 2016
- Publisher :
- Springer New York, 2016.
-
Abstract
- In eukaryotes, RNA polymerase II-driven transcription and processing results in the formation of numerous mRNA 3' isoforms that for any given gene may differ from one another by as little as a single nucleotide. These 3' isoforms can vary in physical properties that may affect their function and stability. Here, we outline a systematic framework to measure individual mRNA 3' isoform half-lives on a genome-wide level in S. cerevisiae. Our approach utilizes the Anchor-Away system to sequester RNA polymerase II (Pol II) in the cytoplasm followed by direct single-molecule RNA sequencing to generate a highly detailed view of 3' isoform stability under most physiological conditions without many of the adverse effects associated with commonly used alternative approaches.
Details
- ISBN :
- 978-1-4939-3066-1
- ISBNs :
- 9781493930661
- Database :
- OpenAIRE
- Journal :
- Methods in Molecular Biology ISBN: 9781493930661
- Accession number :
- edsair.doi...........57e2ca83216900976ba084f2065561a6
- Full Text :
- https://doi.org/10.1007/978-1-4939-3067-8_20