Abstract
AbstractMap order, orientation, and gap or overlap distance of closely linked DNA probes may be determined using fluorescent hybridization to decondensed DNA. The linear arrangement of released chromatin fibers not only simplifies the task of gene ordering, but also provides higher resolution with probes separated by greater distances than can be achieved in FISH with intact interphase nuclei. The first basic protocol of this unit describes an alkaline lysis procedure for generating free chromatin from cultured cells for FISH analysis. A support protocol describes an empirical approach to optimize conditions for preparation of free chromatin. An provides a method for producing free chromatin from cultured lymphocytes with drug treatment. The second basic protocol, high‐resolution FISH mapping with free chromatin, is a modification of the method used for FISH mapping of interphase nuclei.
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