Tritiated Thymidine Incorporation Assay, Using Tissue Explants

Tue May 2 17:00:41 EST 2000


We are interested in finding a protocol and/or technical tips for
precipitation/isolation of incorporated counts into genomic DNA of tissue
explants.  We have thus far solubilized the tissues to completion using
proteinase K; now I wondering if we should use TCA, ethanol or some other
method to precipitate the labelled DNA, and how we would go about isolating
it, i.e., centrifugation or filter-absorption(?).  Also, taking one step
back, if there are any protocols on organ-culture tritiated thymidine assays
in general, we would appreciate hearing of them.  Thanks.

Paul Fanning, Ph.D.
Dept. of Orthopaedic Research/MGRJ1124
Jackson Bldg./1102
Massachusetts General Hospital
55 Fruit St.
Boston, MA 02115      

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