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Determining if an mRNA is a Substrate of Nonsense-Mediated mRNA Decay in Saccharomyces cerevisiae.
Umeå University, Faculty of Science and Technology, Department of Molecular Biology (Faculty of Science and Technology). (Marcus Johansson)
2017 (English)In: Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029, Vol. 1507, 169-177 p.Article in journal (Refereed) Published
Abstract [en]

Nonsense-mediated mRNA decay (NMD) is a conserved eukaryotic quality control mechanism which triggers decay of mRNAs harboring premature translation termination codons. In this chapter, I describe methods for monitoring the influence of NMD on mRNA abundance and decay rates in Saccharomyces cerevisiae. The descriptions include detailed methods for growing yeast cells, total RNA isolation, and Northern blotting. Although the chapter focuses on NMD, the methods can be easily adapted to assess the effect of other mRNA decay pathways.

Place, publisher, year, edition, pages
2017. Vol. 1507, 169-177 p.
Keyword [en]
Half-lives, NMD, Northern blotting, RNA isolation, Yeast, mRNA decay, mRNA levels
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-135240DOI: 10.1007/978-1-4939-6518-2_13PubMedID: 27832540OAI: oai:DiVA.org:umu-135240DiVA: diva2:1098136
Available from: 2017-05-23 Created: 2017-05-23 Last updated: 2017-05-23

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CiteExportLink to record
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