I am currently working in an isothermal amplification technique called PSR ( Polymerase spiral Reaction). I have followed the reaction mixture from previous articles but i am getting primer dimers only in the gel. Can someone please help me out.
If you follow the exact procedure used by previous articles I would suggest looking at your reaction mixture components. Use a fresh set of dNTPS, polymerase and buffer. If this doesn't help, have a look at the starting concentration of your template DNA and perhaps try out a gradient PCR. I have no experience with a PSR PCR, but I would use this approach for a regular PCR.
I have optimized my PCR and it is working perfectly. There are no reports of PSR with my bacteria of interest. That's why i am finding it difficult. Also, the components used were new since we are working for the first time in PSR, we didn't want to add negative factors. So everything is new. And the concentration is prepared correctly.
- Badges
- Science method