In article <dhs3-250794105341 at 184.108.40.206>,
dhs3 at postoffice.cornell.mail.edu (daniel schott) wrote:
> Does anyone have expertise in band-shift (protein-Dna binding) assays? A
> friend of mine is having horrible trouble repeating a series of
> experiments, her samples get stuck in the wells. Due to the nature of the
> experiment, she has to use very low concentrations of DNA and no
> non-specific DNA competitor. She doesn't think the problem is aggregation.
> Is the purity of the acrylimide absolutely vital for this stuff to work?
> Reply to bd14 at cornell.edu, and I'll pass on suggestions. Thanks!
a definative answer to this depends a lot on the source of the protein. We
use bandshift a lot for retic lysates and cell extracts, in these cases the
absence of non-specific competitor gives exactly the result you describe,
everything stuck in the wells. The usual purpose of bandshift is to show a
specific interaction between DNA and protein so why can't she use