DNA comparison

Ryan Duggan rcduggan at midway.uchicago.edu
Tue Mar 5 17:56:08 EST 2002



We're trying to estimate the amount of DNA in the cells of a crustacean
species by comparing the PI fluorescence of it to PI fluorescence of
Drosophila cells.  We are having problems both with the sample prep of
drosophila (embryos or whole flies) and crustacean (we basically grind them
up) and staining of the cells.  Our major problem lies in the fact that we
get so much debris that it is difficult to find populations of cells (or
nuclei).  If anyone has done anything similar to this, your experiences
would be helpful and if anyone has specific experience measuring DNA in
drosophila or crustacean-type species, that would be greatly appreciated
also.  We are using a BD LSR to analyze; 20ug/mL PI staining after
detergent permeabilization (not fixed), and varying concentrations of cells
(1x10^5 cells to 5x10^6 cells)


Thank You,

Ryan




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