I started work with Dream taq polymerase and its buffer but I have primer dimers. For solving this problem I tried gradient PCR from 60 to 66 °C. Is this problem from my primers or my agent? Could you recommend some solution?
Primer dimers are a problem with your primer design. If you are not limited to those exact sequences, I would strongly suggest you change them. Otherwise, your best option is to try touchdown PCR.
Primer dimers are a problem with your primer design. If you are not limited to those exact sequences, I would strongly suggest you change them. Otherwise, your best option is to try touchdown PCR.
I agree with Matthew. I have worked with dreamtaq with no problem. If you have other primers test them to exclude any protocol or buffer problem. But try to modify your primers either in binding position or in length if they must be located at that position.
If you have expected band and primer dimer, please use less primer (sometimes 1/10 is enough)
If you don't have expected band but only primer dimer, please redesign primers and test the reaction system by using other primers which can work well in other systems previously.
I've used Dream taq polymerase (Thermoscientific) for years and always had better results with a (NH4)2SO4, 0.1% Tween 20 buffer than with KCl, provided with this Dream Taq.
Agree with comments above, I also used lower primer conc. Of course, you should check and maybe redesign your primers; you can use Primer3 and OligoAnalyzer tools.
- Badges
- Science topic