Mini Agarose Gel Problem

Ken Howe howe at biology.ucsc.edu
Tue Nov 18 11:42:45 EST 1997


Hey,
you may be pouring your agarose too hot.  What happens is that the top
(ie. exposed to air) part of your gel becomes slightly more concentrated
due to evaporation as your gel cools.  Cooling your agarose by swirling
(in cold water) the container you've melted the agarose in before pouring
it should minimize this effect.  We've found that our larger gels have a
more pronounced slanting effect in the middle of the gel.  Even if we
subsequently cut the gel into smaller segments, the regions corresponding
to what was once the middle still slant more, supporting the idea that
this has to do with cooling and not differences in current strength.  Of
course, you'll have to confirm this one way or the other for your boxes.

 On Tue, 18 Nov
1997, Prince LoLo wrote:

> Date: Tue, 18 Nov 1997 06:33:35 GMT
> From: Prince LoLo <g2bfunky at ix.netcom.com>
> To: methods at net.bio.net
> Subject: Mini Agarose Gel Problem
> 
> I am having problem resolving DNA on a mini (5cm x 8cm x 3mm) agarose
> gel. 1X TBE.  The bands are slanted, such that the DNA closer to the
> top gel layer travels slower than DNA closer to the bottom layer.
> Observing the gel from the side, the bands shape like a
> right-side-left "C".  Slowing the running condition to 1 V/cm or
> reducing the sample did not remedy the situation.
> 
> Can anyone speculate the probable cause of the problem?
> 
> To reply via e-mail, please subsitute "nospam" with "ix".
> 
> 
> 




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