Abstract
In vivo metabolic pulse labeling is a classical approach to assess production and processing of ribosomal RNA (rRNA). However, conventional labeling techniques can be indirect and require work with radioactivity. Here, we describe in detail a protocol for in vivo metabolic labeling, purification, and readout of nascent rRNA by 4-thiouridine (4sU). We propose 4sU labeling as standard nonradioactive technique for the analysis of rRNA metabolism during ribosome biogenesis.
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Acknowledgements
We thank Lars Doelken, Bernd Raedle, and Lisa Marcinowski for help with experimental design and technical support. This work was supported by the Deutsche Forschungsgemeinschaft (SFB684 and SFB1064).
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Burger, K., Eick, D. (2016). A Nonradioactive Assay to Measure Production and Processing of Ribosomal RNA by 4sU-Tagging. In: Németh, A. (eds) The Nucleolus. Methods in Molecular Biology, vol 1455. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3792-9_10
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DOI: https://doi.org/10.1007/978-1-4939-3792-9_10
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