Soaking in nucleic acids into a PAGE gel

Karl Fischer tyr-2 at bones.biochem.ualberta.ca
Thu Jul 12 12:17:06 EST 2001


> I'm not sure it will be possible. You can elute oligos out of acrylamide
> so you don't want to assay using a small polynucleotide. If it can go in
> it can also come out during washes, hence why denatured/activated
> xsomal DNA was used originally.
>
> Why do you want to omit the DNA?

Thanks for the response Duncan,

My colleague is concerned that the proteins of interest are showing
anomalous migration patterns potentially due to interaction with the
polynucleotide during the electrophoresis. By way of example, an enzyme
with an amino terminus deletion shows a slower migration in the activity
gel than the parent enzyme even though the deletant is only 2/3 as large.
He was hoping that leaving out the template from the matrix followed by
template soak-in would give a better sizing comparison.

Karl




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