Sequencing gels

pmiguel at bilbo.bio.purdue.edu pmiguel at bilbo.bio.purdue.edu
Wed Apr 20 14:19:47 EST 1994


In article <joiner-150494155409 at joiner.med.upenn.edu>, joiner at pobox.upenn.edu (Michael Joiner) writes:
>Can anyone tell me how to minimize leakage between adjacent wells when
>using polyacrilamide gels with sharkstooth combs? We seem to have tried
>every combination of prewarming with/without comb, new vs old combs, and
>insertion depth. Whether or not we get leakage still seems to be a random
>occurrence! Does everybody have this trouble?
>
>Mike

  In my experience leakage is a big problem if I don't use spacers and 
combs from the same set.  That is, we have both biorad (double fine) and 
IBI combs and spacers for both.  I make sure I use the biorad spacers with 
the double fine combs as they will have been machined from the same piece 
of plastic and have the same thickness.  Then I clamp the combs (flat side 
down, of course) in after pouring the gel.  
  I've actually had problems at times getting the double fine teeth 
inserted if I don't do it this way.  Then I end up chipping my plates up 
trying to pry them apart a little to fit the teeth in.

Phillip



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