From: Michael Ormerod (Michael_Ormerod@compuserve.com)
Date: Tue Dec 22 1998 - 14:39:32 EST
I cannot comment about about primary cultures of mouse keratinocytes but endoduplication (as described by Derek) is quite common in cell lines - far more common than most cell biologists realise. When doing a DNA analysis is essential to perform a pulse shape analysis to distinguish clumps from single cells. Sometimes you will observe single cells with a DNA content greater than 4n. If you then turn down the gain on the DNA PMT you will usually see a peak at 8n (I.e., endoduplicated cells in G2). You can demonstrate endoduplication very nicely by labelling the cells with an antibody to cyclin B1. There is a nice example I obtined from Jim Jaccoberger on my CD-ROM. Michael Ormerod 34 Wray Park Road Reigate RH2 ODE Telephone: voice & FAX: (0)1737 241726 Web site: http://ourworld.compuserve.com/homepages/Michael_Ormerod
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