Mysterious probes do not bind southern

Francois Nantel Nantel at titus.u-strasbg.fr
Wed Jul 26 10:58:57 EST 1995


In article <DCBMA7.A78 at uns.bris.ac.uk> Harry Witchel,
Harry.Witchel at bristol.ac.uk writes:
>Blotting failure takes one of two forms: either the probe fails to bind 
>anything except for the lambda hind3 control (in which case I lower the 
>temperature), or the probe binds as a smear to all DNA, in a pattern 
>identical to what I see in ethidium (in which case I raise the 
>temperature).  With each separate probe, I try a variety of temperatures 
>and either get nothing or everything.  Have I overlooked any 
>possibilities?  My protocols follow.

<Stuff deleted>

The same thing happened to me a while ago. It turned out that the
fragment I used as a probe was contaminated with the DNA standard. It was
purified from an agarose gel and the standard was in the well next to it.
I redid the digestion and the migration (with not standard this time) and
it worked fine.

Francois
nantel at titus.u-strasbg.fr

IGBMC
Strasbourg, France



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