Sequence gel fixing query...

bugg at mbcf.stjude.org bugg at mbcf.stjude.org
Tue Jun 9 18:08:44 EST 1992


In article <1992Jun4.194330.3389 at magnus.acs.ohio-state.edu>, rrumpf at magnus.acs.ohio-state.edu (Robert Rumpf) writes:
 
> So you don't dry the gels at all?  How much is your signal quenched (if at 
> all), and do you wrap both sides of the gel (front & back)?
> 
> In a related area, if one doesn't remove urea from gels with a methanol/acetic 
> acid preparation, is the urea sucked into the vacuum pump with any stray water 
> vapor, and can this harm the pump?
> 
> Bob

To answer your questions:  I use P-32 and I don't dry the gels at all.  I have
never used S-35.  I don't believe the signal from P-32 gels is quenched at all
but I have never tried an experiment to verify this.  We usually leave the gels
in a film casette with one intensifier screen at -70C for 8 hours to overnight
and we get good reads.  We only wrap the side of the film (or paper) with the
gel on it, but we fold under the edges onto the other side.  Then, we put the
wrapped gel in the casette with the completely covered side up and put the film
on top of it.  It also helps to include one of those otherwise worthless pieces
of yellow-orange paper that Kodak includes in the film boxes between the
wrapped gel and the film you are exposing - prevents Saran-Zap and humidity (we
live in the South) from causing marks on your film.  Good luck!
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Barbara Bugg, M.S.  bugg at mbcf.stjude.org       "I refuse to have a battle of
St. Jude Children's Research Hospital          wits with an unarmed person."
Memphis,  TN  USA
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