Abstract
Determination of RNA stability is one of the basic issues addressed in studies of RNA metabolism. In a standard approach used for RNA half-life measurement synthesis of RNA is inhibited and then the steady-state level of RNA is quantified and used for calculations. Here, we present an optimized protocol for mitochondrial RNA stability studies without perturbation of transcription and present results produced for the mitochondrial CytB messenger RNA. This method was originally described for nuclear transcripts and involves metabolic labeling of RNA with 4-thiouridine.
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Acknowledgements
The authors thank Aleksander Chlebowski for his help with the manuscript. Authors are supported by the Ministry of Science and Higher Education of Poland [0542/IP1/2011/71]. LSB is the recipient of the Stipend for Young Researchers from the Foundation for Polish Science. RJS is the recipient of a TEAM scholarship from the Foundation for Polish Science (TEAM/2008-2/1) and a scholarship for outstanding young scientists from the Polish Ministry of Science and Higher Education.
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Borowski, L.S., Szczesny, R.J. (2014). Measurement of Mitochondrial RNA Stability by Metabolic Labeling of Transcripts with 4-Thiouridine. In: Rorbach, J., Bobrowicz, A. (eds) Polyadenylation. Methods in Molecular Biology, vol 1125. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-971-0_22
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DOI: https://doi.org/10.1007/978-1-62703-971-0_22
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-970-3
Online ISBN: 978-1-62703-971-0
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