Setting Up PCR Reactions Using Phusion PCR Master Mix


Carefully mic and centrifuge all tubes before opening to improve recovery. PCR reactions should be set up on ice.

Component Volume/
50 µl reaction
20 µl reaction
Final Conc.
H2O add to 50 µl add to 20 µl  
2x Phusion Master Mix 25 µl 10 µl 1X
Primer A * x µl x µl 0.5 µM
Primer B * x µl x µl 0.5 µM
Ttemplate DNA x µl xµl  
(DMSO **, optional) (1.5 µl) (0.6 µl) (3%)

* The recommendation for final primer concentration is 0.5 μM, but it can be varied in a range of 0.2-1.0 μM if needed.
** Addition of DMSO is recommended for GC-rich amplicons. DMSO is not recommended for amplicons with very low GC% or amplicons that are >20kb.