NONRADIOACTIVE SEQUENCING GELS

Jerry Kropp jkropp at itsa.ucsf.edu
Mon Jan 27 18:48:28 EST 1997


In article <1997Jan24.220750.8637 at mcrcr6>, oddouc01 at mcrcr6.med.nyu.edu.
(oddouc01) wrote:

> We have been blotting sequencing gels prepared with biotinylated
> primers and find that subseqent detection reagents do not permeate all
> layers well when the blot is coiled in the hybridization tube.  We have
> tried various meshes (Denville) rolled up with the blot "jelly roll"
> style which makes it either worse or adds to background.  Has anyone
> found a method for hybridizing oversize membranes in hybridization
> tubes?
With apologies for a partial answer:
I gave up on the hyb. bottles. Now using the industrial equivalent of a
Seal-a-Meal bag and sealer from National Bag--800-247-6000 (no relation).
For agitation I use a variable speed platform rocker. That's a bit of an
investment in hardware, tho' much cheaper than a Hyb. oven. It works.

This response is gonna cost you, viz., do you (like me) find your bands
are not as sharp as when you used, say S-35? I figure it's a function of
the transfer step.
Hope this helps,
Jerry

-- 
"If it ain't broke, it needs more features"-plagerized



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