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    Synchronizing Progression of Schizosaccharomyces pombe Cells from G2 through Repeated Rounds of Mitosis and S Phase with cdc25-22 Arrest Release.

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    Authors
    Hagan, Iain M
    Grallert, Agnes
    Simanis, V
    Affiliation
    CRUK Cell Division Group, Cancer Research UK Manchester Institute, University of Manchester, Manchester M20 4BX,
    Issue Date
    2016
    
    Metadata
    Show full item record
    Abstract
    Transient inactivation of the cdc25(+) gene product by manipulation of the culture temperature for cdc25-22 cells is the most commonly exploited approach to mitotic synchronization in fission yeast. Because Cdc25 removes the inhibitory phosphate placed on Cdk1 by Wee1, inactivation of Cdc25 arrests cells at the G2/M boundary. Incubation at the restrictive temperature of 36°C for just over one generation time forces all cells in the culture to accumulate at the G2/M boundary. Restoration of Cdc25 function via a return to the permissive temperature or chemical inhibition of Wee1 activity at 36°C can then promote a highly synchronous wave of cell division throughout the culture. These approaches can be performed on any scale and thus support simultaneous assessment of numerous events within a single culture. After describing this simple and widely applicable procedure, we discuss frequently overlooked issues that can have a considerable impact on the interpretation of data from cdc25-22 induction-synchronized cultures.
    Citation
    Synchronizing Progression of Schizosaccharomyces pombe Cells from G2 through Repeated Rounds of Mitosis and S Phase with cdc25-22 Arrest Release. 2016, 2016 (8):pdb.prot091264 Cold Spring Harb Protoc
    Journal
    Cold Spring Harbor Protocols
    URI
    http://hdl.handle.net/10541/619921
    DOI
    10.1101/pdb.prot091264
    PubMed ID
    27480720
    Type
    Article
    Language
    en
    ISSN
    1559-6095
    ae974a485f413a2113503eed53cd6c53
    10.1101/pdb.prot091264
    Scopus Count
    Collections
    All Paterson Institute for Cancer Research

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