i am using the proper protocol for electrophoresis, but the glasses have been broken down and the buffer does heat. I reduced the V, however, it still heats.
I would say that unless you are using a new gel apparatus that you may have something going wrong with the terminals that transduce the electrical current. We use a BIORAD system and one of them would constantly short out, which would turn the machine off by going into an error mode. However, we called them, they replaced the system and no problems since. ALternatively, it is also possible to place a cold freezer pack in with the gel as you run it. This also has the benefit of requiring less SDS running buffer because the freezer pack takes up considerable volume, while also maintaining a lower temperature.
How much is the voltage you set? Normally Voltage should set about the 100V.
If the problem is persistent, set the current rather than voltage.
If the problem is not solve yet, it might be due to faulty in the machine, At this point, I do suggest you to call service center
You can run at constant current. Depending on the size of your gel, you may run it at 12-20mA. Also, as already suggested check the concentration of SDS PAGE buffer you are using, I keep a 10X buffer solution and dilute it to working standard for running. Sometimes it may happen if the buffer is not freshly prepared or may be of a higher concentration than working standard. Also check your running apparatus.
Hope it helps!
boutThanks everyone .
Charles Zogzas: I use same company machine (Biorad) but I changed the machine from the same company and I reduced the voltage to 160 and still same problems with glasses and with the temp. however, the time has been reduced to 1h.
Wai Chin Chong : we were using 250v, 40 mA and we were getting fantastic results.
this problem has been happened suddenly.
Tias Saha: we use same conditions that you are saying, but I am not sure a the running apparatus. do you mean the wires and the machines???
I have three machines, All gave me same problem.
I have checked the buffers of the gels proberly. however, it still same problem.
Regards.
Manaf
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