I am doubtful about this because once the dye comes out of the gel while running, it gets mixed with the running buffer, also sometimes the gel gets over run. So in such cases is it ok to re use the running buffer ?
Firstly the dye shouldnt get dispersed in the running buffer. You will have to ensure the wells are of the same height and that while loading your sample the samples don't flow over the well height. Secondly, dye doesnt affect the running buffer integrity. You will have to check the pH of the running buffer and its sp.conductivity to ensure it is good enough to reuse. Thirdly try filtering the running buffer if it has added on a dye through whatman 41 filter paper. usually the blue dye has more affinity for cellulose rather than the buffer itself. I stand corrected if any part of this communication is erroneous
We routinely re-use the running buffer at-least twice or thrice. The dye should not affect the running of the gel. If the buffer goes bad due to re-using it, often, the current/ voltage does not go up to the desired value. Therefore, the gel does not run at all and you get an error message on the powerpack. So, if this happens to you, you might need to make fresh running buffer. Otherwise, you should be able to re-use the same running buffer twice or thrice safely.
Sometimes, if the buffer goes bad, the gel runs crooked. But this can also happen if the buffer level is low. In this case, if you re-add fresh running buffer, the gel starts running properly again. So, the bottom line is: You can safely re-use the running buffer upto three times. Good luck!
Hi, You can always use the running buffer as many times as you want as the lower tank buffer. For the upper tank, i recommend using fresh buffer each time.
The reason, is, if you gel over runs, then, possibility is that, some of the protein you are working with might enter the buffer, and on reusing it for both upper tank and lower tank, there are possibilities that, your protein resolution may not be as discrete as you expect. So, I recommend you to use fresh upper tank buffer and reuse the lower tank buffer.
thank you all for your prompt reply. I will definitly go thru your suggestions.
thank you all once again
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