glued gels sticking to both plates

Eric Kofoid kofoid at biology.utah.edu
Wed Mar 23 10:27:11 EST 1994


For 13 years, I have glued sequencing gels to their short plates. The only
problems I ever had were initially, when I neglected to clean the excess
glue off the plate with several EtOH wipes. The result was diffusion of the
glue across the gel, adhering it to the long plate. This caused the matrix
to neatly split midway through its plane when the plates were separated.
Not a pleasant sight!

Suddenly, it's happening again! Not across the entire gel surface, but
generally near the spacers, or towards the bottom of the gel, especially in
the corners. Very embarrassing, as I actually *teach* sequencing to people
around here!

Possibly relevant differences: 

     1. Runs have been fairly long lately (8-10 hrs)
     2. Switched to USB's "Glycerol Tolerant Gel Buffer", as I've been
        using their manganese solutions (which work quite well, by the way)
     3. My bottle of 3-(trimethoxysilyl)propyl methacrylate (TSPM) is
        getting pretty old.
     4. I've been using proper wedge spacers from IBI. Previously, I would 
        put an inch long chunk of spacer between the two plates at the top
        and a stack of two between them at the bottom. Capillarity 
        kept the liquid from spilling out the sides.
     5. Some of my plates are getting pretty old.
     6. Some of my plates have been touched by other humans, a nasty 
        condition indeed.

For the record, my glue solution is made by combining 0.1 ml TSPM,
20 ml 95% EtOH and 0.06 ml glacial acetic acid. I store it at room
temperature. Plates are washed with detergent and plastic scouring pad,
rinsed with distilled water followed by 95% EtOH. A single KimWipe is
folded into quarters, half saturated with glue solution, and rubbed
methodically over the entire surface of the short plate. After standing at
room temperature a few minutes, it is washed four times, each time by
spraying with a stream of 95% EtOH onto the surface and wiping it down with
a couple of KimWipes. Long plates are similarly washed and treated with
SigmaCote diluted in chloroform. I'm so finicky that I even change gloves
between types of plates when prepping!

Please, don't tell me to dry down on paper! Glue worked wonderfully for me
for more than a decade and I just want it to work again.

Humble pie and cheers,

Eric.
                                               -- Dept. Biology
                                               -- U. of Utah



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