DNase
Dominique MARIE (marie@sb-roscoff.fr)
Mon, 25 Nov 1996 15:45:25 +0100
Tanks for the responses about DNase treatment, especially for Martin and
Temi. More informations are necessary for a good understanding of my
difficulties. I work with picophytoplankton, and I would like to measure the
RNA content of prochlorophytes. This organism have a small size ranging from
0.4 to 0.6 microns and can be distinghuised from heterotrophic bacteria on
the base of his chlorophyll content. The use of detergents will cause
damages resulting in the loss of the chlorophyll emission and these
prochlorophytes will become undistinghuisable from the bacteriplankton.
The samples are fixed with aldehydes in order to keep the pigment contents
into the cells. Samples are collected during oceanographic cruises, and the
concentrations of picoplanktonic cells are to low and cells are to fragile
to applied physical treatments such as centrifugation, filtrations, that
induce artifacts. How can fixatives interfere with DNase digestion ?
I already have tested the effect of componds including EDTA without
success. I have not tested the effect of DNase on fresh samples because in
my point of view, it's a to large molecule that do not penetrate the membranes.
Have a nice day.
Dominique,
e-mail: marie@sb-roscoff.fr
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