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Transcript Localization in Dictyostelium discoideum Cells by RNA FISH

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Dictyostelium discoideum Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 983))

Abstract

Biogenesis of ribosomal RNA (rRNA) takes place preliminary in the nucleolus of eukaryotic cells, the site of rDNA transcription. Several processing steps of rRNA molecules have been implied to take place in the cytoplasm. To follow these processing events we have adapted protocols for fluorescence in situ hybridization (FISH) for use in Dictyostelium discoideum. We describe methods for the generation of suitable fluorescently labeled probes and the fixation of cells, by which we have localized different precursor and mature rRNA molecules to the nucleolus or the cytoplasm, respectively.

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Acknowledgment

This work was supported by a Heisenberg stipend of the DFG to C.H. (HA 3459/5-2).

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Authors and Affiliations

  1. Heisenberg Research Group Ribogenetics, Technical University of Darmstadt, Darmstadt, Germany

    Patrick Hofmann

  2. Ribogenetics@Biochemistry Laboratory, Jacobs University Bremen, Bremen, Germany

    Janis Kruse & Christian Hammann

Authors
  1. Patrick Hofmann
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  2. Janis Kruse
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  3. Christian Hammann
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Corresponding author

Correspondence to Christian Hammann .

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Editors and Affiliations

  1. , Centre for Biochemistry, University of Cologne, Joseph-Stelzmann-Str. 52, Köln, 50931, Nordrhein-Westfalen, Germany

    Ludwig Eichinger

  2. , Centre for Biomedical Research, University of Hull, Hull, HU6 7RX, Montserrat

    Francisco Rivero

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Hofmann, P., Kruse, J., Hammann, C. (2013). Transcript Localization in Dictyostelium discoideum Cells by RNA FISH. In: Eichinger, L., Rivero, F. (eds) Dictyostelium discoideum Protocols. Methods in Molecular Biology, vol 983. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-302-2_17

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  • DOI: https://doi.org/10.1007/978-1-62703-302-2_17

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-301-5

  • Online ISBN: 978-1-62703-302-2

  • eBook Packages: Springer Protocols

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