Table 3.

Phusion HF PCR conditions.

	Temperature	Duration
Initial denaturation	98 °C	30 s
35 amplification cycles	98 °C	5–10 s
	55 °C (adjust according to primer properties)	10–30 s
	72 °C	15–30 s/kb
Final extension	72 °C	10 min
Hold for storage	4 °C	∞