TAE, TBE vs TGE - what is the difference? (Mar/03/2006 )

Hi.
I'm doing EMSA, and my shifted bands are always bent in a very ugly way.

Sombody recommend I change the chamber bfr from TBE to TGE, but I doubt this helps. Becuase I'm not sure of the function of glycine or boric acid.

Anybody knows the difference among TAE, TBE, and TGE in electrophoresis?

Thanks in advance.

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Good question. someone told me that their function is the same. al6though I noticed that I cant reuse TAE as many times as I reuse TBE.
Sure this didnt help

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hi
differences of these 3 acids came from pKa. Acetate has a pKa of 4.6.
boric acid has 3 pKa : 9.27 12.7 13.8
glycine is a weak base... and it's pKb is 4.22 so can adjust the pH...
for complete refs, go to http://www.chemtutor.com/acid.htm part buffers

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Thanks.
Basic chemistry is important and difficult sometimes.

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Can you explain that more in detail? What is the influence of the different pKa/pKb of a running buffer on EMSA experiment? In my case, I used TBE for EMSA from the begining. I found the bends were not in a sharp way. I changed running buffer to TGE, bends became sharp and beautiful even I had run them in the same percentage gel with TBE.

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I thought TBE is usually used for better resolution of running RNA sample or small fragment samples

I dont know about the TGE. What is it for?

I reuse TAE after about 4-5 runs. After that, it will become cloudy and affect the efficiency of DNA migration. How about you all? How many times can you reuse your buffer?

Hehe... I am still new in molecular biology. Just giving my 2 cents.

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I think TBE and TGE can be reused for many times ( maybe over 5 times) at least in my case no matter DNA electrophoresis or EMSA. But I don't know why TGE made bends more sharp and clear.

In general DNA electrophoresis, I used TBE or TAE depending on my lab cause TAE is much cheaper. I only used TGE in EMSA.

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