Maxipreps and columns (or not?)
Richard P. Grant
see_sig at cmtech.co.delete.uk
Wed Jun 10 10:45:01 EST 1998
I would submit that the problem with transfection-quality DNA is not to do
with the DNA per se, which is what one sees on the gel, rather that there
would be contaminants that are not detectable by EtBr.
DNA is DNA is DNA, but other components in the prep may well upset your
cells, therefore leading to loss of transfection ability. Indeed, this
also affects PCR, sequencing, restriction . . .
In article <6lm7t8$j5o at mserv1.dl.ac.uk>, wgschech at med.uni-tuebingen.de wrote:
: Hi all.
: Just run a quality control on a column maxiprep (NucleoBond from
: macherey-nagel). I noticed no difference in the look of the gel bands
: comparing the material I obtained after lysis and neutralization
: (KOAc step, i.e. that what I apply to the column) and the eluate from
: the column.
: So, I just wonder if all this fuss really is necessary (for doing
: transfections) or if simply a direct iPropOH precipitation would do
: it, too.
: Has anyone yet tried the transfection quality of not column or CsCl
: purified plasmids?
: The E. coli host I'm using is XL1-blue
Richard P. Grant MA DPhil | rgrant at cmtech.co.uk
Senior R&D Scientist | work: http://www.cmtech.co.uk/
Cambridge Molecular | home: http://www.avnet.co.uk/adastra
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