On Jul 16, 2007, at 12:01 PM, bputcha wrote:
I am trying to crystallize a protein-DNA complex. I purify the
protein finally
using gel filtration. I purchase
single stranded complementary oligos (desalting from idtdna.com),
mix them up
How can I purify the duplex DNA further?
in my experience, you need to purify your dna. one way to do it is
to do a trityl on
synthesis, so you can first separate the full length dna from the
rest by reverse phase
hplc, then remove the trityl group, and reverse phase purify again
the clean, full length
dna. this is a general description of a method that i believe comes
from a. aggarwal,
you could check for papers, or the site: http://
atlas.physbio.mssm.edu/~aagroup/
it has been a while, and there may be better methods by now, but
recent references
from groups like the aggarwal group that are doing protein-nucleic
acid complexes
would be a place to start.
best,
--knut
Boston Biomedical Research Institute... Today's Research for
Tomorrow's Health.
Please visit us at www.bbri.org
