plasmid size & transcfection

Carsten Hohoff hohoff at uni-muenster.de
Mon Sep 28 14:27:24 EST 1998


Is the  uptake of CaP/DNA complexes a function of 'moles' or of merely
'mass' ??

I wonder if I have promoter constructs ranging from .2 up to 2.4 kb and take
the same mass amount, the smallest construct will be used in a higher molar
concentration that the largest construct (and thus may yield highest
reporter gene concentrations).

The same problem occurs when I optimize the amount of pSVbeta-gal: i.e., 6.5
ug are O.K., even 3.25 is sufficient for detection of beta-gal  protein. If
I than take 3.25 u g of the different promoter-constructs (fudes to pCAT) I
may get the same problem with normalization, if more moles of vector in the
calcium phosphate transfection solution give more vector taken up and thus
(?) more protein expressed...

Any suggestion is appreciated

Carsten Hohoff

Dept of Biochem
Muenster
Germany 




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