We are doing Western Blots with home made gels. The problem we are having since a couple of months is that the glass cover plates regularly break when gel-electrophoresis is finished.
We use a constant current of 40mA (aprox. 80V).
Is this because of the swelling of the gel or because of the heat building up? Any tips anyone?
I don't know with type of system you use, but try not to overtight the system when you prepare the gel. It is not necessary. Because during polymerization and migration the volume of gel slightly increase.
40mA this is almost boiling, do not exceed 25. The low voltage at this currency suggest 1.5mm spacer (I use 0.75mm) or the buffer composition is wrong. The reason you observe it at the end is probably due to the glycine gradient is leaving the gel in smile shape is causing the tension causing your catastrophe:(
run the gel at 4 degree C or in a cold room, lower the voltage, it will work
Please check your buffer concentration, run the the gel at lower voltage over night, Run the gel in the Air conditioned room
We use self prepared gels as well and even run the electrophoresis at 100 V without any problems. Did you change any component of your western blot system / buffer, since your question suggests that it worked before.
I think it could be the heat causing the problem. If you're re-using the electrophoresis buffer it can generate excessive heat. Use fresh buffer to run the gels under constant voltage settings. Room temperature is enough. No need of 4 degree.
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