Protocol

Cohesin and Condensin

Volume 1515 of the series Methods in Molecular Biology pp 245-255

Date:

A Protocol for Measuring Mitotic Chromosome Condensation Quantitatively in Fission Yeast Cells

  • Christoph SchiklenkAffiliated withCell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL)
  • , Boryana PetrovaAffiliated withCell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL)
  • , Christian H. HaeringAffiliated withCell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL) Email author 

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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.

Key words

Chromosome segregation Genome architecture Mitosis Live cell microscopy Schizosaccharomyces pombe