Reaction setup:

Reagent	Amount
5× Q5 Reaction Buffer	10 μL
10 mM dNTPs	1 μL
10 μM Forward Primer	2.5 μL
10 μM Reverse Primer	2.5 μL
Q5 DNA Polymerase	0.5 μL
Template Plasmid (5 ng)	1.0 μL
Nuclease-Free Water	33.5 μL
Total	50 μL