band separation prob. on agarose gels

EK someone at microsoft.net
Fri Feb 20 03:44:28 EST 2004


Do you by any chance boil your DNA samples before running on a gel? Some
labs do boil DNA. If so, there is a chance that your particular fragments do
not anneal correctly, and rather form predominantly 2 species, one of them
running coincidentally with the band you want to get rid of. That might
indeed had to do with the reagents (loading buffer?) you used. Could that be
true? Otherwise, I am same flabbergasted as you are.
Emir

"Kyle Legate" <legatek at hotmail.com> wrote in message
news:cdea459ff6bffb2bd990677c829976ab at news.teranews.com...
> Kyle Legate wrote:
> > It all started when I wasn't getting any colonies on my plates, so I
> > checked the purity of my starting material...
> >
> <snip long description of purifying two fragments from one band...>
>
> I received some good suggestions into how to fix this problem, but in the
> end nothing worked so I threw out and remade all reagents except for the
> restriction enzymes. That seems to have fixed the problem. Somehow it
almost
> always comes down to this solution, for specific reasons I'll never know.
>
>





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