Buffer BPTE for DNA electrophoresis - (Feb/02/2007 )

Does anyone has experience with buffer BPTE for DNA PAGE?
For some reason, we changed the bufferring condition for our DNA PAGE. But at such pH (~6.5), we could not get good resolution.

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just for check you made 10 x BPTE electrophoresis buffer by adding 100 mM PIPES free acid, 300 mM Bis-Tris free base, 10 mM EDTA, pH 8,0. Final pH 6,5. is that so?

could you please send us your PAGEs photo?

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Thank you!

Yes, I prepare the 10 X PBTE as you state.

Finally, I found the reason. We use APS and TEMED as initiators. At pH 6.5, the effect of TEMED is weakened and our TEMED is oxidized. Then I increased the amount of TEMED, and get excellent resolution.

The uploaded file states almost all factors affecting the quality of acrylamide gel. Very helpful!

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two standard samples before and after I increased the amount of TEMED( attachment)
Both 20% acrylamide gel, same APS, only difference is the TEMED

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