I am trying to perform emulsion PCR with the EURX micellula DNA emulsion and purification kit. After the PCR, a precipitate is formed at the bottom of the PCR tubes, which I assume is the micelles breaking down. I have tried different polymerases ( Q5® Hot Start High-Fidelity DNA Polymerase and DreamTaq polymerase ) and different denaturing temperatures (82-95 C) but the same problem persists.
Does anyone has insights into the issue?
The emulsion doesn't looked properly formed to me, its not very milky in the before picture. The "precipitate" you describe appears be the emulsion with the poorly mixed oil phase floating on top. The first tube looks as though the aqueous phase separated completely at the bottom, then emulsion layer, and finally oily phase on top.
Are you sure Dreamtaq buffer doesn't contain detergents?
Were you able to solve the problem? I'm experiencing something similar.
- Badges
- Science method