Abstract
The native template of all eukaryotic nuclear RNA polymerases is chromatin. To understand how transcription occurs in vivo, it is important to define the chromatin environment of transcribing RNA Pols. Here, we describe a method used to characterize the distribution and the protein environment of RNA Pol I on ribosomal DNA during transcription in the yeast S. cerevisiae. The method is based on conventional chromatin immunoprecipitation and we propose quality control analyses at different steps of the procedure. Finally, the obtained samples are a useful source for downstream analyses by semiquantitative mass spectrometry or quantitative PCR.
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Acknowledgments
We thank all members of the “Institute für Biochemie III” for their support and for helpful discussions. The help of Prof. Dr. Rainer Deutzmann and Eduard Hochmuth in establishing downstream mass spectrometric analysis is gratefully acknowledged. We thank Prof. Dr. Herbert Tschochner, Dr. Philipp Milkereit, and Dr. Joachim Griesenbeck for their support and critical reading of the manuscript.
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Bruckmann, A., Linnemann, J., Perez-Fernandez, J. (2016). Purification of RNA Polymerase I-Associated Chromatin from Yeast Cells. 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_16
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DOI: https://doi.org/10.1007/978-1-4939-3792-9_16
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-3792-9
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