Abstract
Even though the formation of compact cylindrical chromosomes early during mitosis or meiosis is a prerequisite for the successful segregation of eukaryotic genomes, little is known about the molecular basis of this chromosome condensation process. Here, we describe in detail the protocol for a quantitative chromosome condensation assay in fission yeast cells, which is based on precise time-resolved measurements of the distances between two fluorescently labeled positions on the same chromosome. In combination with an automated computational analysis pipeline, this assay enables the study of various candidate proteins for their roles in regulating genome topology during cell divisions.
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Acknowledgements
We are grateful to Kota Miura (CMCI, EMBL Heidelberg) for advice and developing the Fiji software plugin for data analysis and to the EMBL Advanced Light Microscopy Facility (ALMF) for assistance. Work in the authors’ laboratory is funded by EMBL and grants HA5853/1-2 and HA5853/2-1 from the German Research Foundation.
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Schiklenk, C., Petrova, B., Haering, C.H. (2017). A Protocol for Measuring Mitotic Chromosome Condensation Quantitatively in Fission Yeast Cells. In: Yokomori, K., Shirahige, K. (eds) Cohesin and Condensin. Methods in Molecular Biology, vol 1515. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6545-8_15
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DOI: https://doi.org/10.1007/978-1-4939-6545-8_15
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6543-4
Online ISBN: 978-1-4939-6545-8
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