Protocol

Mitosis

Volume 545 of the series Methods in Molecular Biology pp 243-258

Date:

The Spindle Checkpoint: Assays for the Analysis of Spindle Checkpoint Arrest and Recovery

  • Josefin FerniusAffiliated withWellcome Trust Centre for Cell Biology, University of Edinburgh
  • , Kevin G. HardwickAffiliated withWellcome Trust Centre for Cell Biology, University of Edinburgh

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Abstract

The spindle checkpoint is a surveillance mechanism that ensures the fidelity of chromosome segregation by inhibiting anaphase onset until all chromosomes have established stable bipolar attachments. Here we describe a number of protocols that can be used to assay the ability of budding and fission yeast cells to (1) establish and maintain a spindle checkpoint arrest, and (2) segregate chromosomes efficiently upon recovery from mitotic arrest. We focus on experimental detail of the budding yeast protocols, but also point out important differences between budding and fission yeast assays.

Key words

Checkpoint segregation recovery biorientation