Hi ruheng,
Since you synthesized the oligos, you probably already know if there is
any residual salt or buffer in your oligos. I don't know if that
caused the problem. Sometimes people purify and desalt the oligos
before the annealing step.
Joe
ruheng wrote:
Dear CCP4bbers,
I am no
Ru Heng,
It is commonly helpful to combine your protein and DNA under
dilute conditions and then concentrate the complex. Combining
concentrated DNA and protein together has a very good chance of
precipitating in my experience. I completely agree that trying
different buffer conditions
Couple of things, Ru Heng.
1. What buffer conditions is your protein in? Is it similar to the
buffer you describe as using to dissolve your DNA in? In general, you
can even get away with dissolving and annealing the oligos in just
Tris etc.
2. Play with buffer conditions, particularly NaCl
Dear CCP4bbers,
I am now working on a DNA binding protein and the purity of the protein is
quite good, however the results of DLS showed that the protein aggregates
terribly in quite a lot of different buffer conditions I tried and still no
crystals can be obtained. So I am going to co-c
