Component	25 μl Reaction	Final concentration
2X Q5 Mastermix (NEB	12.5 μl	1X
10 μM Forward Primer	1.25 μl	0.5 μM
10 μM Reverse Primer	1.25 μl	0.5 μM
Template DNA (1ng/pl)	1.0 μl	0.04ng/ul
Nuclease-Free Water	9.0 μl	Up to 25 μl
Step	Temperature	Time
Initial Denaturation	98°C	30s
	98°C	10s
14 Cycles	67°C	10s
	72°C	15s
Final Extension	72°C	2mins
Hold	4°C	Hold