TBE vs TAE for agarose gel electrophoresis - (Aug/06/2006 )
hi!!
i just want to noe if anyone can share with me the recipe for 40XTAE buffer with me. i juz want to compare with mine coz after gel electhrophoresis, my gel n the tank became hotter than when i use TBE buffer before. juz wanna check if i did messed up in the recipe. thanks!!
I have used TAE atleast three times and i had no probs.. TBE has better resolution on DNA gels, and for DNA it is better to use 1X conc. for protein 0.5 X is okay
good luck
she
I would just like to know, is there any major difference between TBE buffer and TAE buffer for agarose gel electrophoresis. Plus, is there any different between 0.5x TBE and 1.0x TBE in the results?
Thank you.
Hi,
Yes there is a difference between TBE and TAE that is their electroconductivity
In case of agarose gel electrophoresis it is better to use TBE
0.5X TBE has half the empirical amount of Tris, boric acid and EDTA ahere as in case of the 1.0X TBE it contains the empirical amount of the buffer constituents.
I have seen from the net somewhere that its better to avoid TBE in extraction of band if you are to use in cloning or synthesis due to the borate salt and use TAE instead. I notice this when I use platinum taq for amplification and the gel I run tapered at the end but not present when I rerun using TAE gel and buffer.
but we use most of the time TBE....
in our lab, we use 0.25X TBE in our tanks and gels (2-2.5% agarose).
our products are around 200 kb (+/- 100)
we change (refresh) these every monday.
we run approx 25 gels 1hr each at 200V (constant voltage) per week in each of our tanks.
we have no problems, though we do at times need to topup the buffer due to eveaporation.
hope that helps.
I use TAE all this while. everyone in my lab use TAE... we reuse many many many times, for whole week... still OK.
I think I agree with friley, in my lab also we use TAE for cloning whereas in other neighbouring labs I've seen that people use TBE instead of TAE, I dint know why but now I guess bcoz they do it for polymorphism. [
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I partialy solved my problem with increased heating of the TAE buffer, because I run it in the fridge
I can use a slight higher voltage and the bands look better, IMO.
I've noticed that if I allow my gels to set in the fridge (and get really cold) then the bands run sharper. It's particularly good for running restriction digests because they have a higher salt content so they run hotter and produce blurry bands (especially at 250v+)
I use .5X TAE all the time and have gotten excellent results from PCR/RT-PCR products. For better signal I sometimes add 2~4uL EtBr to the + end buffer solution in the apparatus.