11/21/2023 0 Comments Actin western blot size![]() Basically the wires, the buffer, and the gel itself have charge carrying properties and the conductivity of all three are suspect here. No current means there is a break somewhere or too much resistance. Turns out the wires actually provide noticeable resistance if they are wearing out. I use an old power source and gel tank so I get this problem sometimes too. Check any seals/gaskets on the tanks too - if they were all bought at the same time, they might all be wearing out at the same time. ![]() If they don’t fit tightly enough, the buffer could be leaking out of the middle chamber over the course of the run. If this is suddenly happening when it never used to, and happening with multiple different tanks, I’d also check and make sure there wasn’t a change in the type of cassettes your lab orders. Running the gels with the tanks sitting in some ice works well to limit evaporation, if that ends up being the problem. If this is the issue, the gel should start moving again if you just top up the middle buffer chamber, no need to replace all the buffer. u/coolhmk definitely check whether your buffer level is getting low in the middle chamber. It also fits with the issue being fixed by changing the buffer (because OP would re-fill the setup to the correct level). The buffer level dropping in the middle chamber makes perfect sense, and would lead to gels that run fine for a while, then stop when the buffer level gets low. So many people here are suggesting OP forgot to remove the tape seal or made their running buffer wrong, or has a faulty machine, but OP says the gel runs completely fine up to a certain point and then stops, which doesn’t really fit any of those suggestions. I’d guess it’s this as well - evaporating or possibly leaking.
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