Hi all, I proceed with nucleic acid gel electrophoresis recently several times, but a wried phenomenon is they always melted after running over 30min.
My gel concentration is 1% percentage Agarose, parameters are 90V, 400mA. If the running time is 20min, the gel will safe and sound, but once I prolong the running time, the gel will be melted in the TAE buffer.
Does anyone meet with such situations?
Dear Wnxin
did you dissolve the agarose in water or TAE buffer?
Manuele
As Manuele Martinelli mentions, the melting has to be the result of Joule heating due to lack of buffer in the conductive material, this case the gel. Even low-melting point agarose won't melt in properly buffered conditions. Double check that you are making your gel in the same buffer conditions as your running buffer. Also, a very minor but often overlooked point is to measure the weight of the gel/buffer solution before heating. It is typical to loose as much as 10% of the solution volume during heating/microwaving and this should be replaced back with DI water for optimal results. Good luck!
Thomas J Esparza Hi Thomas, thanks for your answer, I will try your suggestion again and see the result. Wenxin
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