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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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Western blot separating gel problem
Posted by: ncl08 (IP Hidden, New member, 1)
Date: February 20, 2008 08:09AM
Hello I am an undergraduate project student. I am trying to cast a 12% separating gel using the BioRad Mini-protean II equipment. I am pouring the solution (which has been degassed and contains 50ul APS and 5ul TEMED) between the plates using an automatic pipette up to the fill line 1cm below where the bottom of the lanes will be. I then overlay the solution with water saturated butanol, being careful not to mix the two solutions, and I leave to polymerise for approx 1hr. When I return to the gel after this time, somehow the level of the separating gel has decreased and fallen well below the original fill line. I have checked for leaks in the bottom of the plate and both myself and my supervising phd student are at a miss to explain what is happening. Does anyone have any idea?
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