Aaron,
I have had this problem before with 2.6 mm ID IEF tubes and
also with DNA sequencing gels. Applied Biosystems calls the
phenomena "electroendoosmosis." Something to do with impurities on
the glass and the inability of some detergents/cleaning procedures to
remove it. ABI recommended using ethanolic base followed by
ethanolic acid treatment of the glass with a final rinse in Milli-Q
water. I have used that procedure with great success. Also what
works is a good soak in RBS-35 (Pierce Chemical Co.) in hot water
followed by thorough rinse with Milli-Q water. If all other
parameters of your IEF system are normal this should solve the
problem.
Good Luck,
Brian Hampton
>hello all,
>
>i'm running mini 2-d gels using biorad's mini IEF cell and slab
>system. my acrylamide polymerized approximately 2/3 to 3/4 of the
>way up the tube. however, after a run of 20000 vhr, i find the
>tubes have expanded, some even out of the gasket. as it was
>impossible to remove the gasket without breaking the gel, i lost
>some mass of my tube gels.
>
>so there's my problem. my question is, how do i avoid this? we
>think it may be due to thermal expansion of the gel, so we are
>considering running the gels at 4 C. obviously, we could also cast
>less gel in the tube, but we were wondering if anyone else has
>encountered this phenomenon and what you did to avoid/solve it.
>thanks!
>
>Aaron Downs
>
>_____________________________________________________________
>Let the world know you are a lab rat!
>
>FREE yourname@thelabrat.com email addresses available at
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Project Leader, Hematopoiesis Department
American Red Cross, Holland Laboratory
15601 Crabbs Branch Way
Rockville, MD 20855
hampton@usa.redcross.org
Voice: 301-738-0814
Fax: 301-738-0811
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