FAQ: I am having trouble amplifying a template that is longer than 5kb. How can I optimize my product yield using Phusion® High-Fidelity DNA Polymerase?

* Use more template. Sample concentration may be too low.
* Template DNA may be damaged. Use carefully purified template.
* Optimize enzyme concentration by testing a titration of enzyme in the reaction (0.25-2 units/50μl reactions)
* Increase number of cycles.
* If HF buffer has failed, try using GC buffer. GC buffer allows better amplification of GC rich and longer templates, but slightly lowers fidelity.
* Lengthen extension time.