FAQ: How should I set up a PCR reaction using Q5® Hot Start 2X Master Mix?

The general guidelines for a 50 µl reaction are: 

0.5 μM each primer
2-50 pg plasmid or 50-500 ng genomic template
1X Q5i Hot Start High-Fidelity 2X Master Mix
Denature at 98°C
Extend 20 s/kb at 72°C 

Reactions can be set up at room temperature