I ran the gel at 60 volts, and then 100 volts. By the time of completion the entire box was warm . I had it filled in and around the dam with about a liter of running buffer.
Hello David.
Clean the apparatus by soaking them in detergent completely immediately after every use, this includes glass plates, sponge pads, plastic holder, too. Then tap water, H2O, and dry in the air for next use.
There are so many remnants dried at each use. Particles, dusts, dried salts and even microbes, too. All these would make resistance higher under electric current. It will get the apparatus hot.
Prepared transfer buffer should be filtered through two layers of Whatman filters to remove any possible particles.
Perform the electric transfer in cold room at 4C or in an ice-filled Styrofoam box for watch.
Best wishes.
We normally place ice packs that fit just nicely in the transfer tank as we mostly run 2 transfer reactions in tanks which can actually fit 4 transfer reactions. A little warmth is to be expected as explained above, due to ohmic heating and waste heat, but as long as your gel shape is not affected, your reaction should run fine. I recall one instance when I tried running a gel at 250 V for 30 min (as suggested by one of my supervisors to save time) and I ended up having a half-melted gel which was ruined.
Please be sure that you are using a lower concentration of your transfer buffer (1X) and it's better to keep ice packs inside the apparatus or you can also keep the same in refrigerator during the transfer process.
Hi David:
You can use MilliQ water intead of dH2O to prepare all the nwestern blot buffers. That will decrease the resistance to electric current in your system.
Good luck!!!!!!
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