another question on DNA-quatitizing

pathos pathos at freerealtime.com
Wed Feb 9 19:06:25 EST 2000


In article <4.2.0.58.20000209183506.009692c0 at marlin.bio.umass.edu>,
rashie at bio.umass.edu (rashie athukorala) wrote:

> something else to keep in mind with respect to the point about fluorescing 
> with equal intensity is fact that the 100 bp band, since it has travelled a 
> longer distance in the gel, would be more diffuse than the 3000 bp 
> band.  one would then have to think in terms of a dilution series to build 
> a saturation curve from which you can quantify your unknowns.

Under the assumption that the latitudinal diffusion is similar for both
species which would be valid if the length of time for the gel is under an
hour or so, the integration of the the  entire peak (not the max value) 
should yield good numbers regardless of the "band broadening" that will
occur due to the differences in the londitudinal diffusion which is
accentuated by the different distances traveled by the to species.  Thus a
dillution series is not needed.

-- 
Peter Pediaditakis
University of Pittsburgh
Dept. of Pathology




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