UV damage and cloning, was Re: aberrant DNA migration on an agarose gel

Rob Jordan rjordan at u.washington.edu
Tue Aug 1 18:43:35 EST 2000


> An alternative (and I think easier) approach is to go ahead and run the
gel
> with EtBr and then make sure to visualize with a UV source that won't
> damage the DNA, e.g. a long-wave hand held UV lamp is what I use.  I've
> used this approach for years without any problems cloning.  Of course I'm
> still careful to try to minimize the UV exposure, but I think the
> EtBr-stained DNA can still take a lot of long wave UV without damage.

What I've done (after someone suggested it here) is to put 1mmol/L cytidine
in the running and staining buffer. I don't do that much cloning so I don't
know how much it actually helps in my case, but it's very simple. Here's the
ref:

Grundemann and Schomig, Protection of DNA during preparative agarose gel
electrophoresis against damage induced by ultraviolet light. Biotechniques
21:989-903 (November 1996).

Rob


--
Robert Jordan
Radiation Biology Laboratory
University of Washington Medical Center
Box 356069
Seattle, WA 98105








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